Cleaning compositions comprising enzymes

ABSTRACT

The present invention relates to compositions, methods and use of a mixture of enzymes having DNase activity and an enzyme having hexosaminidase activity.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a 35 U.S.C. 371 national application ofPCT/EP2017/063489 filed June 2, 2017, which claims priority or thebenefit under 35 U.S.C. 119 of Denmark application PA 2016 00328 filedJun. 3, 2016, the contents of which are fully incorporated herein byreference.

REFERENCE TO A SEQUENCE LISTING

This application contains a Sequence Listing in computer readable form,which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to cleaning (detergent) compositionscomprising an enzyme having deoxyribonuclease (DNase) activity and anenzyme having hexosaminidase activity, a method for laundering a textileand the use of an enzyme having DNase activity together with an enzymehaving hexosaminidase activity. The invention further relates to use ofcleaning compositions comprising an enzyme having deoxyribonuclease(DNase) activity and an enzyme having hexosaminidase activity incleaning processes.

BACKGROUND OF INVENTION

Some bacteria are capable of adhering to textiles (clothing) and forminga biofilm on the textile. Biofilm may be present on laundry items, suchas fabrics, hard surfaces, such as dish wash utensils, dish washers andwashing machines where they may cause malodor, which is difficult toremove even after wash. Biofilm may also make laundry items sticky andsoil adheres to the sticky areas. Further, when very dirty laundry itemsare washed together with less dirty laundry items the dirt present inthe wash liquor tend to stick to the biofilm.

Enzymes comprising hexosaminidase activity include chitinase and the useof such enzymes is described in WO9850512 (Proctor and Gamble). Enzymeshaving hexosaminidase activity include Dispersins such as Dispersin B(DspB), which as described is β-N-acetylglucosamininidases belonging tothe Glycoside Hydrolase 20 family. WO04061117 A2 (Kane Biotech INC)describe compositions comprising DspB for reducing and preventingbiofilm caused by poly-N-acetylglucosamine-producing bacteria anddescribes the use of the compositions comprising DspB forreduction/removing biofilm on medical devices and for wound care.

WO 2015/155350 (Novozymes A/S) discloses the use of a polypeptide havingDNase activity for preventing, reducing or removing a biofilm componente.g. DNA from an item, wherein the polypeptide is obtained from a fungalsource, such as A. oryzae and the item is a textile.

SUMMARY OF THE INVENTION

The present invention relates to a composition comprising an enzymehaving DNase activity, an enzyme having hexosaminidase activity and anadjunct ingredient, wherein the hexosaminidase is selected from thegroup consisting of the DspB clade, the Curtobacterium clade and theTerribacillus clade. The invention further relates to the use of a suchcomposition for deep cleaning of an item, wherein the item is a hardsurface or a textile.

The invention further relates to a method for laundering an itemcomprising the steps of:

-   -   a. Exposing an item to a wash liquor comprising (i) an enzyme        having DNase activity and an enzyme having hexosaminidase        activity or (ii) a composition according to any of claims 1-18;    -   b. Completing at least one wash cycle; and    -   c. Optionally rinsing the item,

wherein the item is a textile.

The invention further relates to the use of a combination of an enzymehaving DNase activity and an enzyme having hexosaminidase activity fordeep cleaning an item, wherein the item is a textile or a hard surface.

Definitions

DNase (deoxyribonuclease): The term “DNase” means a polypeptide or anenzyme with DNase activity that catalyzes the hydrolytic cleavage ofphosphodiester linkages in the DNA backbone, thus degrading DNA. Forpurposes of the present invention, DNase activity is determinedaccording to the procedure described in the Assay II or Assay IIa. Inone aspect, the DNase according to the present invention have at least20%, e.g., at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 95%, or at least 100% of the DNaseactivity of the mature polypeptide of SEQ ID NO: 2. In one aspect theDNase of the present invention has at least 60%, e.g., at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, or at least 100% sequence identity to the amino acid sequencehaving SEQ ID NO: 2.

Dispersin: The term “dispersin” and the abbreviation “Dsp” means apolypeptide having hexosaminidase activity, EC 3.2.1.—that catalyzes thehydrolysis of β-1,6-glycosidic linkages of N-acetyl-glucosamine polymers(poly-N-acetylglucosamine) found e.g. in biofilm. Hexosaminidase: Theterm “hexosaminidases” means a polypeptide having hexosaminidaseactivity (hexosaminidases), and includes EC 3.2.1. e.g. that catalyzesthe hydrolysis of N-acetyl-D-hexosamine or N-acetyl-glucosamine polymersfound e.g. in biofilm. The term includes dispersins and includespolypeptides having N-acetylglucosaminidase activity andβ-N-acetylglucosamininidase activity. The term “polypeptide havinghexosaminidase activity” may be used interchangeably with the termhexosaminidases and similar the term “polypeptide havingβ-N-acetylglucosaminidase activity” may be used interchangeably with theterm β-N-acetylglucosamininidases. For purposes of the presentinvention, hexosaminidase activity is determined according to theprocedure described in Assay I. In one aspect, the polypeptides of thepresent invention have at least 20%, e.g., at least 40%, at least 50%,at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, orat least 100% of the hexosaminidase activity of the mature polypeptideof SEQ ID NO: 14.

Biofilm: A biofilm may be produced by any group of microorganisms inwhich cells stick to each other or stick to a surface, such as atextile, dishware or hard surface or another kind of surface. Theseadherent cells are frequently embedded within a self-produced matrix ofextracellular polymeric substance (EPS). Biofilm EPS is a polymericconglomeration generally composed of extracellular DNA, proteins, andpolysaccharides. Biofilms may form on living or non-living surfaces. Themicrobial cells growing in a biofilm are physiologically distinct fromplanktonic cells of the same organism, which, by contrast, aresingle-cells that may float or swim in a liquid medium. Bacteria livingin a biofilm usually have significantly different properties fromplanktonic bacteria of the same species, as the dense and protectedenvironment of the film allows them to cooperate and interact in variousways. One benefit of this environment for the microorganisms isincreased resistance to detergents and antibiotics, as the denseextracellular matrix and the outer layer of cells protect the interiorof the community. On laundry and hard surfaces biofilm producingbacteria can be found among the following species; Acinetobacter sp.,Aeromicrobium sp., Brevundimonas sp., Microbacterium sp., Micrococcusluteus, Pseudomonas sp., Staphylococcus epidermidis, andStenotrophomonas sp. On hard surfaces biofilm producing bacteria mayinclude but are not limited to the following species: Acinetobacter sp.,Aeromicrobium sp., Brevundimonas sp., Microbacterium sp., Micrococcusluteus, Pseudomonas sp., Staphylococcus epidermidis, Staphylococcusaureus and Stenotrophomonas sp. A wide range of bacterial and fungalmicroorganisms have been found to produce PNAG or PNAG-like surfacepolysaccharides, including but not limited to Bacillus subtillis,Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis,Pseudomonas fluorescens, Yersinia pestis, Aggregatibacteractinomycetemcomitans, Streptococcus pyogenes, Streptococcusdysgalactiae (group C strep), Enterococcus faecalis, Listeriamonocytogenes, Clostridium difficile, Mycobacterium tuberculosis,Mycobacterium smegmatis; Neisseria meningitides, Neisseria gonorrhea,nontypable Haemophilus influenzae, Haemophilus ducreyi, Helicobacterpylori, Campylobacter jejuni; Citrobacter rodentium, Salmonella entericaserovars typhi, Salmonella typhimurium, Candida albicans, Aspergillusflavus, Fusarium solani, and Cryptococcus neoformans.

Extracellular DNA (eDNA) is a common matrix component in microbialbiofilms and has been identified in, but not limited to, Acinetobacterbaumannii, Actinobacillus actinomycetemcomitans, Bdellovibriobacterivorous, Bordetella pertussis, Bordetella bronchiseptica,Campylobacter jejuni, Comamonas denitrificans, Escherichia coli,Haemophilus influenza, Klebsiella pneumoniae, Neisseria meningitides,Pseudomonas aeruginosa, Shewanella oneidensis, Vibrio cholera,Gram-positive bacteria, Bacillus licheniformis, Bacillus subtilis,Enterococcus faecalis, Listeria monocytogenes, Micrococcus luteus,Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcushaemolyticus, Streptococcus anginosus, Streptococcus constellatus,Streptococcus salivarius, Staphylococcus lugdunesis, Streptococcusintermedius, Streptococcus intermedius, Streptococcus mutans,Streptococcus pneumoniae, Streptococcus pyogenes, Aspergillus fumigatusand Candida albicans.

Most biofilms comprises biofilm or EPS from bacteria of many differentspecies and are thus “poly-cultural”.

Clade a group of polypeptides clustered together based on homologousfeatures traced to a common ancestor. Polypeptide clades can bevisualized as phylogenetic trees and a clade is a group of polypeptidesthat consists of a common ancestor and all its lineal descendants e.g.the Terribacillus clade or clade of Terribacillus is a group of enzymesall related to the same ancestor and share common properties.

Coding sequence: The term “coding sequence” means a polynucleotide whichdirectly specifies the amino acid sequence of a polypeptide. Theboundaries of the coding sequence are generally determined by an openreading frame, which begins with a start codon such as ATG, GTG, or TTGand ends with a stop codon such as TAA, TAG, or TGA. The coding sequencemay be a genomic DNA, cDNA, synthetic DNA, or a combination thereof.

Cleaning adjunct ingredient: The cleaning adjunct ingredient is aningredient which is different from the enzymes comprised in the cleaningcomposition of this invention. The term includes the term “cleaningcomponent” and is defined herein to mean the types of components whichcan be used in cleaning compositions. Examples of cleaning componentsare alkalis, surfactants, hydrotropes, builders, co-builders, chelatorsor chelating agents, bleaching system or bleach components, polymers,fabric hueing agents, fabric conditioners, foam boosters, sudssuppressors, dispersants, dye transfer inhibitors, fluorescent whiteningagents, perfume, optical brighteners, bactericides, fungicides, soilsuspending agents, soil release polymers, anti-redeposition agents,enzyme inhibitors or stabilizers, enzyme activators, antioxidants andsolubilizers.

Cleaning composition: The term “cleaning composition” refers tocompositions that find use in the removal of undesired compounds fromtextiles to be cleaned. The term includes detergent compositions. Thecleaning composition may be used to e.g. clean textiles for bothhousehold cleaning and industrial cleaning. The terms encompass anymaterials/compounds selected for the cleaning composition desired andthe form of the product (e.g., liquid, gel, powder, granulate, paste, orspray compositions) and includes, but is not limited to, detergentcompositions (e.g., liquid and/or solid laundry detergents and finefabric detergents; fabric fresheners; fabric softeners; and textile andlaundry pre-spotters/pretreatment). In addition to containing theenzymes of the invention, the cleaning composition may contain one ormore additional enzymes (such as proteases, amylases, lipases,cutinases, cellulases, endoglucanases, xyloglucanases, pectinases,pectin lyases, xanthanases, peroxidaes, haloperoxygenases, catalases andmannanases, or any mixture thereof), and/or cleaning components such assurfactants, builders, chelators or chelating agents, bleach system orbleach components, polymers, fabric conditioners, foam boosters, sudssuppressors, dyes, perfume, tannish inhibitors, optical brighteners,bactericides, fungicides, soil suspending agents, anti-corrosion agents,enzyme inhibitors or stabilizers, enzyme activators, transferase(s),hydrolytic enzymes, oxido reductases, bluing agents and fluorescentdyes, antioxidants, and solubilizers.

Deep cleaning: By the term “deep cleaning” is meant reduction or removalof components of biofilm, such as EPS or parts hereof, polysaccharides,PNAG (poly-N-acetylglucosamine), proteins, DNA, soil or other componentspresent in the biofilm.

Enzyme Detergency Benefit: The term “enzyme detergency benefit” isdefined herein as the advantageous effect an enzyme may add to adetergent compared to the same detergent without the enzyme. Importantdetergency benefits which can be provided by enzymes are stain removalwith no or very little visible soils after washing and/or cleaning,prevention or reduction of redeposition of soils released in the washingprocess (an effect that also is termed anti-redeposition), restoringfully or partly the whiteness of textiles which originally were whitebut after repeated use and wash have obtained a greyish or yellowishappearance (an effect that also is termed whitening). Textile carebenefits, which are not directly related to catalytic stain removal orprevention of redeposition of soils, are also important for enzymedetergency benefits. Examples of such textile care benefits areprevention or reduction of dye transfer from one fabric to anotherfabric or another part of the same fabric (an effect that is also termeddye transfer inhibition or anti-backstaining), removal of protruding orbroken fibers from a fabric surface to decrease pilling tendencies orremove already existing pills or fuzz (an effect that also is termedanti-pilling), improvement of the fabric-softness, colour clarificationof the fabric and removal of particulate soils which are trapped in thefibers of the fabric or garment. Enzymatic bleaching is a further enzymedetergency benefit where the catalytic activity generally is used tocatalyze the formation of bleaching components such as hydrogen peroxideor other peroxides.

Fungal: In the context of the present invention the term “fungal” inrelation to polypeptide (such as an enzyme, e.g. a DNase) refers to apolypeptide encoded by and thus directly derivable from the genome of afungus, where such fungus has not been genetically modified to encodesaid polypeptide, e.g. by introducing the encoding sequence in thegenome by recombinant DNA technology. In the context of the presentinvention, the term “fungal DNase” or “polypeptide having DNase activityobtained from a fungal source” or “polypeptide is of fungal origin” thusrefers to a DNase encoded by and thus directly derivable from the genomeof a fungal species, where the fungal species has not been subjected toa genetic modification introducing recombinant DNA encoding said DNase.Thus, the nucleotide sequence encoding the fungal polypeptide havingDNase activity is a sequence naturally in the genetic background of afungal species. The fungal polypeptide having DNase activity encoding bysuch sequence may also be referred to a wildtype DNase. In one aspect ofthe invention the enzyme having DNase activity is substantiallyhomologous to a fungal DNase. In the context of the present invention,the term “substantially homologous” denotes a polypeptide having DNaseactivity which is at least 80%, preferably at least 85%, more preferablyat least 90%, more preferably at least 95%, even more preferably atleast 96%, 97%, 98%, and most preferably at least 99% identical to theamino acid sequence of SEQ ID NO 2.

Isolated: The term “isolated” means a substance in a form or environmentthat does not occur in nature. Non-limiting examples of isolatedsubstances include (1) any non-naturally occurring substance, (2) anysubstance including, but not limited to, any enzyme, variant, nucleicacid, protein, peptide or cofactor, that is at least partially removedfrom one or more or all of the naturally occurring constituents withwhich it is associated in nature; (3) any substance modified by the handof man relative to that substance found in nature; or (4) any substancemodified by increasing the amount of the substance relative to othercomponents with which it is naturally associated (e.g., recombinantproduction in a host cell; multiple copies of a gene encoding thesubstance; and use of a stronger promoter than the promoter naturallyassociated with the gene encoding the substance). An isolated substancemay be present in a fermentation broth sample; e.g. a host cell may begenetically modified to express the enzymes of the invention. Thefermentation broth from that host cell will comprise the isolatedpolypeptide.

Laundering: The term “laundering” relates to both household launderingand industrial laundering and means the process of treating textileswith a solution containing a cleaning or detergent composition of thepresent invention. The laundering process can for example be carried outusing e.g. a household or an industrial washing machine or can becarried out by hand.

Malodor: By the term “malodor” is meant an odor which is not desired onclean items. The cleaned item should smell fresh and clean withoutmalodors adhered to the item. One example of malodor is compounds withan unpleasant smell, which may be produced by microorganisms. Anotherexample is unpleasant smells can be sweat or body odor adhered to anitem which has been in contact with human or animal. Another example ofmalodor can be the odor from spices, which sticks to items such as curryor other exotic spices which smell strongly.

Sequence identity: The relatedness between two amino acid sequences orbetween two nucleotide sequences is described by the parameter “sequenceidentity”.

For purposes of the present invention, the sequence identity between twoamino acid sequences is determined using the Needleman-Wunsch algorithm(Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implementedin the Needle program of the EMBOSS package (EMBOSS: The EuropeanMolecular Biology Open Software Suite, Rice et al., 2000, Trends Genet.16: 276-277), preferably version 5.0.0 or later. The parameters used aregap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62(EMBOSS version of BLOSUM62) substitution matrix. The output of Needlelabeled “longest identity” (obtained using the -nobrief option) is usedas the percent identity and is calculated as follows:(Identical Residues×100)/(Length of Alignment−Total Number of Gaps inAlignment)

Textile: The term “textile” means any textile material including yarns,yarn intermediates, fibers, non-woven materials, natural materials,synthetic materials, and any other textile material, fabrics made ofthese materials and products made from fabrics (e.g., garments and otherarticles). The textile or fabric may be in the form of knits, wovens,denims, non-wovens, felts, yarns, and towelling. The textile may becellulose based such as natural cellulosics, including cotton,flax/linen, jute, ramie, sisal or coir or manmade cellulosics (e.g.originating from wood pulp) including viscose/rayon, cellulose acetatefibers (tricell), lyocell or blends thereof. The textile or fabric mayalso be non-cellulose based such as natural polyamides including wool,camel, cashmere, mohair, rabbit and silk or synthetic polymers such asnylon, aramid, polyester, acrylic, polypropylene and spandex/elastane,or blends thereof as well as blends of cellulose based and non-cellulosebased fibers. Examples of blends are blends of cotton and/orrayon/viscose with one or more companion material such as wool,synthetic fiber (e.g. polyamide fiber, acrylic fiber, polyester fiber,polyvinyl chloride fiber, polyurethane fiber, polyurea fiber, aramidfiber), and/or cellulose-containing fiber (e.g. rayon/viscose, ramie,flax/linen, jute, cellulose acetate fiber, lyocell). Fabric may beconventional washable laundry, for example stained household laundry.When the term fabric or garment is used, it is intended to include thebroader term textiles as well. In the context of the present invention,the term “textile” also covers fabrics.

Variant: The term “variant” means a polypeptide/enzyme having the sameactivity as the parent enzyme comprising an alteration, i.e., asubstitution, insertion, and/or deletion, at one or more positions. Asubstitution means replacement of the amino acid occupying a positionwith a different amino acid; a deletion means removal of the amino acidoccupying a position; and an insertion means adding an amino acidadjacent to and immediately following the amino acid occupying aposition. In the context of the present invention, a variant may be avariant of an identified DNase that has the enzymatic activity of theparent, i.e. the capacity of catalyzing the hydrolytic cleavage ofphosphodiester linkages in the DNA backbone (deoxyribonucleaseactivity). In one embodiment, the deoxyribonuclease activity of thevariant is increased with reference to the parent DNase, e.g. thepolypeptide of SEQ ID NO: 2.

In the context of the present invention, a variant may also be a variantof an identified hexosaminidase that has the enzymatic activity of theparent, i.e. the capacity of catalyzing the hydrolysis ofβ-1,6-glycosidic linkages of N-acetyl-glucosamine polymers(hexosaminidase activity). In one embodiment, the hexosaminidaseactivity of the variant is increased with reference to the parenthexosaminidase, e.g. the polypeptide of SEQ ID NO: 14.

Wash cycle: The term “wash cycle” is defined herein as a washingoperation wherein textiles are immersed in the wash liquor, mechanicalaction of some kind is applied to the textile in order to release stainsand to facilitate flow of wash liquor in and out of the textile andfinally the superfluous wash liquor is removed. After one or more washcycles, the textile is generally rinsed and dried.

Wash liquor: The term “wash liquor” is intended to mean the solution ormixture of water and at least a surfactant, optionally including otherdetergent components e.g. enzymes other than the enzyme having DNaseactivity, and which is used for laundering textiles.

Wash performance: One way of measuring the wash performance is the Deltaenzyme performance value (ΔRem enzyme value): The term “Delta enzymeremission value” is defined herein as the result of a reflectance orremission measurement at 460 nm. The remission value of a swatch ismeasured and compared to a swatch of similar colour as background,preferably a swatch from a repetition wash, with a swatch representingeach type being measured before wash. The Delta enzyme remission is theremission value of the swatch washed in detergent with an enzyme presentminus the remission value of a similar swatch washed in a detergentwithout enzyme present.

Another way of measuring the wash performance is by use of the Colordifference (L value): A Lab color space is a color-opponent space withdimension L for lightness. The L value, L*, represents the darkest blackat L*=0, and the brightest white at L*=100. In the context of thepresent invention L value is also referred to as color difference.

Whiteness: The term “whiteness” is defined herein as e.g. greying oryellowing e.g. textiles. Greying and yellowing can be due to soilredeposition, body soils, colouring from e.g. iron and copper ions ordye transfer.

DETAILED DESCRIPTION OF THE INVENTION

It was surprisingly found that washing textiles with an enzyme havingDNase activity (DNase) in combination with an enzyme havinghexosaminidase activity (Hexosaminidase) gives a surprisingly goodresult with regard to deep cleaning e.g., reduction/removing of biofilmcomponents, maintaining whiteness and reducing redeposition of soil andmalodor. A wide range of bacterial and fungal microorganisms have beenfound to produce PNAG or PNAG-like surface polysaccharides and mostbiofilms will comprise some amount of PNAG or PNAG-like surfacepolysaccharides since many different ubiquitous bacteria possess theability to produce PNAG. DNA is also present in many biofilms.Therefore, DNA and PNAG is usually present in a biofilm includingbiofilms associated with cleaning such as laundry, washing machines,hard surfaces e.g. utensils etc. The present invention has thereforewide applications. One aspect of the invention relates to the use of acomposition e.g. cleaning composition comprising a combination of anenzyme having DNase activity (DNase) and an enzyme having hexosaminidaseactivity (Hexosaminidase) for deep cleaning of an item e.g.reducing/removing biofilm components, wherein the biofilm componentscomprises PNAG and DNA. The biofilm components may be associated withtextiles, such as clothing, washing machines, hard surfaces e.g. in ADW.

In one aspect, the biofilm or EPS producing strain is Staphylococcusepidermidis. In one aspect the biofilm or EPS producing strain isPseudomonas alcaliphila or Pseudomonas fluorescens. In one aspect, thebiofilm or EPS producing strain is Staphylococcus aureus. As described,adding a combination of enzymes having hexosaminidase activity e.g.Dispersins together with a DNase e.g. from A. Oryzae, such as a DNasewith SEQ ID NO 2 or having at least 60% sequence identity hereto, isadvantageous as the complex construction of biofilms gives rise tohidden pockets of PNAG and DNA inside the biofilm, pockets that are notavailable when only one enzyme is added. Applying the compositioncomprising the enzyme combination of the present invention providesbetter deep cleaning effect than by adding the single specificities.Surprisingly, it was found that adding a composition comprising theenzyme having hexosaminidase activity and the enzyme having DNaseactivity at the same time provides a synergistic effect e.g. as eachenzymes' specific action releases more biofilm component than if theenzymes were added separately, see e.g. example 2 and 3. Thus, oneaspect of the invention relates to the use of a composition comprising ahexosaminidase and a DNase for deep cleaning of an item, wherein asynergistic deep cleaning effect is obtained.

Further, this synergistic effect was shown in a detergent on e.g. ontextiles such as laundry items. In one aspect of the invention thecombination of the enzyme having hexosaminidase activity e.g. thedispersins and the enzyme having DNase activity e.g. the A. oryzae DNaseprovides synergistic deep cleaning effect in a detergent and is thusparticularly useful in cleaning applications such as laundry and hardsurface cleaning e.g. ADW (automatic dish washing).

One aspect relates to a method for laundering an item comprising thesteps of:

-   -   a) Exposing an item to a wash liquor comprising an enzyme having        DNase activity and enzyme having hexosaminidase activity or a        composition according to the invention;    -   b) Completing at least one wash cycle; and    -   c) Optionally rinsing the item.

One aspect of the invention relates to a method of laundering a textile,comprising the steps of:

-   -   a) Contacting the textile with a wash liquor comprising an        enzyme having DNase activity, an enzyme having hexosaminidase        activity and a surfactant; and    -   b) optionally rinsing the textile,

wherein the enzyme having DNase activity and the hexosaminidase activityhave deep cleaning properties.

In a wash liquor the concentration of each of the DNase andhexosaminidase is preferably at least 0.00001 mg/L or at least 0.0001 orat least 0.001 mg/L of enzyme or at least 0.01, 0.02, 0.05, 0.1, 0.2,0.5, 1, 2, 5, 10 or at least 15 μg active enzyme per liter of washliquor. Optionally the concentration of enzyme in the wash liquor is inthe range of about 0.00002 mg/L to about 2 mg/L, about 0.0002 mg/L toabout 2 mg/L, about 0.002 mg/L to about 2 mg/L, such as about 0.02 mg/Lto about 2 mg/L, such as about 0.2 mg/L to about 2 mg/L or in the rangeof about 0.00001 mg/L to about 1 mg/L, about 0.0001 mg/L to about 1mg/L, about 0.0001 mg/L to about 10 mg/L or in the range of in the rangeof about 0.001 mg/L to about 10 mg/L, or in the range of about 0.01 mg/Lto about 10 mg/L, or in in the range of about 0.1 mg/L to about 10 mg/Lper liter of wash liquor.

In one aspect, the biofilm comprises at least one bacterium having thepolysaccharide poly-N-acetylglucosamine (PNAG) on the outer surface. Inone aspect, such bacteria are Pseudomonas fluorescens. In one aspectsuch bacteria is staphylococcus aureus and the biofilm or EPS comprisesPNAG. Enzymes having hexosaminidase activity, such as Dispersins,hydrolyze β-1,6-glycosidic linkages of N-acetyl-glucosamine polymers,and compositions comprising Dispersin B (DspB) is described inWO200406117. However, to be useful in cleaning processes enzymes need toperform their action under the conditions of cleaning processes such aslaundry, which includes stability in the presence of cleaning componentssuch as surfactants, builders and bleach components. There is noindication in the art of the use of hexosaminidases, such as dispersins,in combination with DNases in cleaning processes such as laundry or indetergent compositions comprising adjuncts e.g. surfactants, buildersand/or bleaches.

Thus, some aspects of the invention relate the use of an enzyme havingDNase activity and an enzyme having hexosaminidase activity in acleaning process. Some aspects of the invention relates to cleaningcompositions comprising a) an enzyme having DNase activity and an enzymehaving hexosaminidase activity and b) at least one surfactant,preferably at least one surfactant selected from the group consisting ofanionic, nonionic and/or cationic surfactants.

Some aspects of the present invention relate to laundry or cleaningcompositions comprising a DNase and a hexosaminidase enzyme, preferablyat a level of from about 0.000001 wt % to about 1 wt %, from about0.0001 wt % to about 1 wt %, from about 0.0002 wt % to about 1 wt %,from about 0.0005 wt % to about 1 wt %, from about 0.001 wt % to about 1wt %, from about 0.002 wt % to about 1 wt %, from about 0.005 wt % toabout 1 wt %, preferably from about 0.01 wt % to about 0.5 wt %,preferably from 0.0002 wt % to about 1 wt % by weight (wt %) of thecomposition. The amounts are wt % per unit active enzyme e.g. from about0.00001 wt % to about 1 wt % of DNase by weight of the composition andfrom about 0.00001 wt % to about 1 wt % of hexosaminidase by weight ofthe composition.

The concentration of the active enzyme having hexosaminidase activity ispreferably at least 0.00001%, preferably at least 0.00002%, preferablyat least 0.0001 wt %, preferably at least 0.0002 wt %, preferably atleast 0.001 wt %, preferably at least 0.002 wt %, preferably at least0.005 wt %, preferably at least 0.01 wt %, preferably at least 0.02 wt%, preferably at least 0.05 wt % preferably at least 0.1 wt % of thetotal detergent concentration.

The concentration of the active enzyme having DNase activity ispreferably at least 0.00001%, preferably at least 0.00002%, preferablyat least 0.0001 wt %, preferably at least 0.0002 wt %, preferably atleast 0.001 wt %, preferably at least 0.002 wt %, preferably at least0.005 wt %, preferably at least 0.01 wt %, preferably at least 0.02 wt%, preferably at least 0.05 wt % preferably at least 0.1 wt % of thetotal detergent concentration.

The amount enzyme may also be in ppm (mg/L) active enzyme protein. Thus,in one aspect the amount of DNase in the composition is at least 0.00001ppm, 0.00002 ppm, 0.00005 ppm, 0.0001 ppm, 0.0002 ppm, 0.0005 ppm, 0.001ppm, 0.002 ppm, 0.005 ppm, 0.01 ppm, 0.02 ppm, 0.05 ppm, 0.1 ppm, 0.2ppm, 0.5 ppm, 1 ppm, 2 ppm, 5 ppm, 10 ppm or at least 20 ppm DNaseenzyme. In one aspect, the amount of DNase in the composition is in therange from about 0.00001 ppm to about 10 ppm, or in the range from about0.0001 ppm to about 2 ppm or in the range from about 0.001 ppm to about2 ppm DNase enzyme.

In one aspect the amount of hexosaminidase in the composition is atleast 0.00001 ppm, 0.00002 ppm, 0.00005 ppm, 0.0001 ppm, 0.0002 ppm,0.0005 ppm, 0.001 ppm, 0.002 ppm, 0.005 ppm, 0.01 ppm, 0.02 ppm, 0.05ppm, 0.1 ppm, 0.2 ppm, 0.5 ppm, 1 ppm, 2 ppm, 5 ppm, 10 ppm or at least20 ppm hexosaminidase enzyme. In one aspect, the amount ofhexosaminidase in the composition is in the range from about 0.00001 ppmto about 10 ppm, or in the range from about 0.0001 ppm to about 2 ppm orin the range from about 0.001 ppm to about 2 ppm hexosaminidase enzyme.

In one aspect, the ratio of DNase to hexosaminidase is 1:2, such as 1:3,such as 1:4, such as 1:5, such as 1:6, such as 1:7, such as 1:8, such as1:9, such as 1:10.

In one aspect, the ratio of hexosaminidase to DNase is 1:2, such as 1:3,such as 1:4, such as 1:5, such as 1:6, such as 1:7, such as 1:8, such as1:9, such as 1:10.

Preferably the amount of hexosaminidase enzyme is higher than the amountof DNase in cleaning composition of the invention.

In the compositions, e.g. cleaning composition, the enzyme levels areexpressed by pure enzyme by weight of the total composition unlessotherwise specified and the adjunct ingredients are expressed by weightof the total composition.

One aspect of the invention relates to a cleaning compositioncomprising:

a) at least 0.0001 wt % e.g. 0.001 wt % of an enzyme having DNaseactivity and at least 0.0001 wt % e.g. 0.001 wt % of an enzyme havinghexosaminidase activity, and

b) from about 0 wt % to about 60 wt % surfactant, preferably from about5 wt % to about 60 wt %, preferably from about 10 wt % to about 60 wt %,preferably from about 15 wt % to about 60 wt %, wherein the surfactantis selected from anionic, nonionic and/or cationic surfactants.

The surfactant may be selected among nonionic, cationic, anionic and/oramphoteric surfactants as described above, preferably anionic ornonionic surfactants but also cationic or amphoteric surfactants may beused. In general, bleach-stable surfactants are preferred. Preferredanionic surfactants are sulphate surfactants and alkyl ether sulphates,especially C9-C15 alcohol ethersulfates, C12-C15 primary alcoholethoxylate, C8-C16 ester sulphates and C10-C14 ester sulphates, such asmono dodecyl ester sulphates. Non-limiting examples of anionicsurfactants include sulfates and sulfonates, in particular linearalkylbenzenesulfonates (LAS), isomers of LAS, branchedalkylbenzenesulfonates (BABS), phenylalkanesulfonates,alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates,alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates,alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcoholsulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates(AES or AEOS or FES, also known as alcohol ethoxysulfates or fattyalcohol ether sulfates), secondary alkanesulfonates (SAS), paraffinsulfonates (PS), ester sulfonates, sulfonated fatty acid glycerolesters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES)including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid,dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives ofamino acids, diesters and monoesters of sulfo-succinic acid or salts offatty acids (soap), and combinations thereof.

The anionic surfactants are preferably added to the detergent in theform of salts. Suitable cations in these salts are alkali metal ions,such as sodium, potassium and lithium and ammonium salts, for example(2-hydroxyethyl) ammonium, bis (2-hydroxyethyl) ammonium and tris(2-hydroxyethyl) ammonium salts.

Non-limiting examples of nonionic surfactants include alcoholethoxylates (AE or AEO), alcohol propoxylates, propoxylated fattyalcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylatedand/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates(APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG),alkoxylated amines, fatty acid monoethanolamides (FAM), fatty aciddiethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM),propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fattyacid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides,GA, or fatty acid glucamides, FAGA), as well as products available underthe trade names SPAN® and TWEEN®, and combinations thereof. Commerciallyavailable nonionic surfactants include Plurafac™, Lutensol™ andPluronic™ range from BASF, Dehypon™ series from Cognis and Genapol™series from Clariant. In one aspect, the surfactant is non-natural i.e.not found in nature.

The cleaning composition of the invention may optionally comprise abuilder and in one aspect of the invention the cleaning composition is alaundry (powder or liquid) or ADW composition comprising:

a) at least 0.0001 wt % e.g. 0.001 wt % of an enzyme having DNaseactivity and at least 0.0001 wt % e.g. 0.001 wt % of an enzyme havinghexosaminidase activity, and optionally

b) from about 0 wt % to about 60 wt % surfactant, preferably from about5 wt % to about 60 wt %, preferably from about 10 wt % to about 60 wt %,preferably from about 15 wt % to about 60 wt %, wherein the surfactantis selected from anionic, nonionic and/or cationic surfactants, andoptionally

c) from about 0 wt % to about 50 wt % of at least one builder,preferably from about 5 wt % to about 50 wt %, preferably from about 10wt % to about 50 wt %, preferably from about 15 wt % to about 50 wt %,wherein the builder preferably is selected from carbonates, zeolites,phosphate builder, calcium sequestering builders or complexing agents.

The builder is preferably selected among phosphates, sodium citratebuilders, sodium carbonate, sodium silicate, and sodium aluminosilicate(zeolite). Suitable builders are alkali metal or ammonium phosphates,polyphosphates, phosphonates, polyphosphonates, carbonates,bicarbonates, borates, citrates, and polycarboxylates. Citrate builders,e.g., citric acid and soluble salts thereof (particularly sodium salt),are polycarboxylate builders. Citrates can be used in combination withzeolite, silicates like the BRITESIL types, and/or layered silicatebuilders. The builder is preferably added in an amount of about 0-65% byweight, such as about 5% to about 50% by weight. In the composition ofthe invention, the level of builder is typically about 40-65% by weight,particularly about 50-65% by weight, particularly from 20% to 50% byweight. The builder and/or co-builder may particularly be a chelatingagent that forms water-soluble complexes with Ca and Mg. Any builderand/or co-builder known in the art for use in cleaning detergents may beutilized. Non-limiting examples of builders include zeolites,diphosphates (pyrophosphates), triphosphates such as sodium triphosphate(STP or STPP), carbonates such as sodium carbonate, soluble silicatessuch as sodium metasilicate, layered silicates (e.g., SKS-6 fromHoechst), and (carboxymethyl)inulin (CMI), and combinations thereof.Further non-limiting examples of builders include citrate, chelatorssuch as aminocarboxylates, aminopolycarboxylates and phosphonates, andalkyl- or alkenylsuccinic acid. Additional specific examples include2,2′,2″-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid(EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid(IDS), ethylenediamine-N,N′-disuccinic acid (EDDS),methylglycine-N,N-diacetic acid (MGDA), glutamic acid-N,N-diacetic acid(GLDA), 1-hydroxyethane-1,1-diphosphonic acid,N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoaceticacid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), asparticacid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA),N-(sulfomethyl)aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid(SEAS), N-(sulfomethylglutamic acid (SMGL), N-(2-sulfoethyl)-glutamicacid (SEGL), N-methyliminodiacetic acid (MIDA), serine-N,N-diacetic acid(SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diaceticacid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilicacid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA) andN′-(2-hydroxyethyl)ethylenediamine-N,N,N′-triacetic acid (HEDTA),diethanolglycine (DEG), and combinations and salts thereof.

Phosphonates suitable for use herein include1-hydroxyethane-1,1-diphosphonic acid (HEDP), ethylenediaminetetrakis(methylenephosphonic acid) (EDTMPA), diethylenetriaminepentakis(methylenephosphonic acid) (DTMPA or DTPMPA or DTPMP), nitrilotris(methylenephosphonic acid) (ATMP or NTMP),2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC), andhexamethylenediaminetetrakis (methylenephosphonic acid) (HDTMP)

The composition of the invention may also contain 0-50% by weight, suchas about 5% to about 30%, of a detergent co-builder. The composition mayinclude a co-builder alone, or in combination with a builder, forexample a zeolite builder. Non-limiting examples of co-builders includehomopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copoly (acrylic acid/maleic acid) (PAA/PMA) orpolyaspartic acid. Further exemplary builders and/or co-builders aredescribed in, e.g., WO 09/102854, U.S. Pat. No. 5,977,053. In onepreferred embodiment, the builder is a non-phosphorus based builder suchas citric acid and/or methylglycine-N, N-diacetic acid (MGDA) and/orglutamic-N, N-diacetic acid (GLDA) and/or salts thereof.

One aspect of the invention relates a composition comprising at leastone enzyme having DNase activity and at least one enzyme havinghexosaminidase activity and a non-phosphate builder selected from citricacid, methyl glycine-N, N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and mixtures thereof.

In one aspect, the composition is cleaning composition, such as alaundry composition or an automatic dish wash composition (ADW)comprising:

a) at least 0.0001 wt % e.g. 0.001 wt % of an enzyme having DNaseactivity and at least 0.0001 wt %, e.g. 0.001 wt % of an enzyme havinghexosaminidase activity, and

b) from about 0 wt % to about 50 wt % of at least one builder,preferably from about 5 wt % to about 50 wt %, preferably from about 10wt % to about 50 wt %, preferably from about 15 wt % to about 50 wt %,wherein the builder preferably is selected from carbonates, zeolites,phosphate builder, calcium sequestering builders or complexing agents,preferably citric acid, methylglycine-N, N-diacetic acid (MGDA) and/orglutamic acid-N, N-diacetic acid (GLDA) and mixtures thereof, andoptionally

c) at least one bleach component.

The composition may contain 0-30% by weight, such as about 1% to about20%, such as about 1% to about 10%, such as about 1% to about 5%, suchas about 10% to about 30%, such as about 5% to about 10% or such asabout 10% to about 20% by weight (wt %) of a bleaching system. Anybleaching system comprising components known in the art for use incleaning detergents may be utilized. Suitable bleaching systemcomponents include sources of hydrogen peroxide; sources of peracids;and bleach catalysts or boosters.

Sources of Hydrogen Peroxide

Suitable sources of hydrogen peroxide are inorganic persalts, includingalkali metal salts such as sodium percarbonate and sodium perborates(usually mono- or tetrahydrate), and hydrogen peroxide-urea (1/1).

Sources of Peracids

Peracids may be (a) incorporated directly as preformed peracids or (b)formed in situ in the wash liquor from hydrogen peroxide and a bleachactivator (perhydrolysis) or (c) formed in situ in the wash liquor fromhydrogen peroxide and a perhydrolase and a suitable substrate for thelatter, e.g., an ester.

a) Suitable preformed peracids include, but are not limited to,peroxycarboxylic acids such as peroxybenzoic acid and itsring-substituted derivatives, peroxy-α-naphthoic acid, peroxyphthalicacid, peroxylauric acid, peroxystearic acid, ε-phthalimidoperoxycaproicacid [phthalimidoperoxyhexanoic acid (PAP)], ando-carboxybenzamidoperoxycaproic acid; aliphatic and aromaticdiperoxydicarboxylic acids such as diperoxydodecanedioic acid,diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassylic acid,2-decyldiperoxybutanedioic acid, and diperoxyphthalic, -isophthalic and-terephthalic acids; perimidic acids; peroxymonosulfuric acid;peroxydisulfuric acid; peroxyphosphoric acid; peroxysilicic acid; andmixtures of said compounds. It is understood that the peracids mentionedmay in some cases be best added as suitable salts, such as alkali metalsalts (e.g., Oxone®) or alkaline earth-metal salts.

b) Suitable bleach activators include those belonging to the class ofesters, amides, imides, nitriles or anhydrides and, where applicable,salts thereof. Suitable examples are tetraacetylethylenediamine (TAED),sodium 4-[(3,5,5-trimethylhexanoyl) oxy] benzene-1-sulfonate (ISONOBS),sodium 4-(dodecanoyloxy) benzene-1-sulfonate (LOBS), sodium4-(decanoyloxy) benzene-1-sulfonate, 4-(decanoyloxy) benzoic acid(DOBA), sodium 4-(nonanoyloxy) benzene-1-sulfonate (NOBS), and/or thosedisclosed in WO98/17767. A family of bleach activators of interest wasdisclosed in EP624154 and particularly preferred in that family isacetyl triethyl citrate (ATC). ATC or a short chain triglyceride liketriacetin has the advantage that they are environmentally friendly.Furthermore, acetyl triethyl citrate and triacetin have good hydrolyticstability in the product upon storage and are efficient bleachactivators. Finally, ATC is multifunctional, as the citrate released inthe perhydrolysis reaction may function as a builder.

Bleach Catalysts and Boosters

The bleaching system may also include a bleach catalyst or booster.

Some non-limiting examples of bleach catalysts that may be used in thecompositions of the present invention include manganese oxalate,manganese acetate, manganese-collagen, cobalt-amine catalysts andmanganese triazacyclononane (MnTACN) catalysts; particularly preferredare complexes of manganese with 1,4,7-trimethyl-1,4,7-triazacyclononane(Me3-TACN) or 1,2,4,7-tetramethyl-1,4,7-triazacyclononane (Me4-TACN), inparticular Me3-TACN, such as the dinuclear manganese complex[(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and[2,2′,2″-nitrilotris(ethane-1,2-diylazanylylidene-κN-methanylylidene)triphenolato-κ3O]manganese(III).The bleach catalysts may also be other metal compounds, such as iron orcobalt complexes.

In some embodiments, where a source of a peracid is included, an organicbleach catalyst or bleach booster may be used having one of thefollowing formulae:

or mixtures thereof; wherein each R1 is independently a branched alkylgroup containing from 9 to 24 carbons or linear alkyl group containingfrom 11 to 24 carbons, preferably each R1 is independently a branchedalkyl group containing from 9 to 18 carbons or linear alkyl groupcontaining from 11 to 18 carbons, more preferably each R1 isindependently selected from the group consisting of 2-propylheptyl,2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl,hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl and isopentadecyl.

Other exemplary bleaching systems are described, e.g., in WO2007/087258,WO2007/087244, WO2007/087259, EP1867708 (Vitamin K) and WO2007/087242.Suitable photobleaches may for example be sulfonated zinc or aluminiumphthalocyanines.

The present invention relates to cleaning compositions e.g. detergentcompositions comprising at least one enzyme having DNase activity and atleast one enzyme having hexosaminidase activity, compositions e.g.detergent compositions, and the use of cleaning e.g. detergentcompositions of the invention for deep cleaning of an item such as atextile.

Accordingly, some aspects of the invention relate to cleaningcompositions comprising:

a) at least 0.0001 wt % e.g. 0.001 wt % of an enzyme having DNaseactivity and at least 0.0001 wt % e.g. 0.001 wt % of an enzyme havinghexosaminidase activity, and optionally

b) from about 0 wt % to about 50 wt % of at least one builder,preferably from about 5 wt % to about 50 wt %, preferably from about 10wt % to about 50 wt %, preferably from about 15 wt % to about 50 wt %,wherein the builder preferably is selected from carbonates, zeolites,phosphate builder, calcium sequestering builders or complexing agents,preferably citric acid, methylglycine-N, N-diacetic acid (MGDA) and/orglutamic acid-N, N-diacetic acid (GLDA) and mixtures thereof, andoptionally

c) from about 0 wt % to about 60 wt % surfactant, preferably from about5 wt % to about 60 wt %, preferably from about 10 wt % to about 60 wt %,preferably from about 15 wt % to about 60 wt %, wherein the surfactantis selected from anionic, nonionic and/or cationic surfactants,preferably selected from anionic surfactants such as LAS, AOS, AEOSand/or nonionic surfactants such as AE or AEO, and optionally

d) from about 0 or 30 wt %, preferably from about 5% to about 30%,preferably from about 10% to about 30% by weight (wt %) of at least onebleach component, preferably selected from percarbonates, persulphatesand peracids, preferably percarbonate or a manganese catalyst,preferably 1,4,7-trimethyl-1,4,7-triazacyclononane or manganese (III)acetate tetrahydrate (MnTACN).

The enzymes having hexosaminidase activity to be used in the cleaningcomposition of the invention, which are useful for deep cleaning ofitems such as hard surfaces, textiles and/or fabric, are selected fromhexosaminidases from the DspB clade, the Curtobacterium clade and theTerribacillus clade. In one aspect, the hexosaminidase is selected fromthe DspB clade. In one aspect the hexosaminidase is selected from any ofthe polypeptides having SEQ ID NOS 3, 4, 5, 6, 7, 8, 9, and 10 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto. The enzymes havinghexosaminidase activity have β-N-acetylglucosamininidase activity,β-1,6-N-acetylglucosaminidase activity and in some aspects, thehexosaminidase activity is β-1,6-N-acetylglucosaminidase activity andthe enzymes of the invention are β-1,6-N-acetylglucosaminidases. Oneaspect of the invention relates to cleaning compositions comprisinghexosaminidases from the DspB clade, except for SEQ ID NO 10. Thus, inone aspect the hexosaminidase is not the hexosaminidase with SEQ ID NO10.

In one aspect, the enzymes having hexosaminidase activity to be used inthe cleaning composition of the invention, which are useful for deepcleaning of items such as hard surfaces, textiles and/or fabric, areselected from hexosaminidases from the Curtobacterium clade. In oneaspect, the hexosaminidase is a polypeptide with SEQ ID NO 11 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto.

In one aspect, the hexosaminidase is a polypeptide with SEQ ID NO 15 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto.

In one aspect, the hexosaminidase is a polypeptide with SEQ ID NO 16 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto.

In one aspect, the hexosaminidase is a polypeptide with SEQ ID NO 17 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto.

In one aspect, the hexosaminidase is a polypeptide with SEQ ID NO 18 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto.

In another aspect, the enzymes having hexosaminidase activity to be usedin the cleaning composition of the invention, which are useful for deepcleaning of items such as hard surfaces, textiles and/or fabric, areselected from hexosaminidases from the Terribacillus clade. In oneaspect the hexosaminidases is selected from any of the polypeptideshaving SEQ ID NOS 12, 13, 14, 19, 20 or polypeptides having at least60%, such as at least 65%, such as at least 70%, such as at least 75%,such as at least 80%, such as at least 85%, or such as at least 90%,such as at least 91%, such as at least 92%, such as at least 93%, suchas at least 94%, such as at least 95%, such as at least 96%, such as atleast 97%, such as at least 98%, such as at least 99% or 100% sequenceidentity hereto. In one aspect, the hexosaminidase is a polypeptide withSEQ ID NO 12 or polypeptides having at least 60%, such as at least 65%,such as at least 70%, such as at least 75%, such as at least 80%, suchas at least 85%, or such as at least 90%, such as at least 91%, such asat least 92%, such as at least 93%, such as at least 94%, such as atleast 95%, such as at least 96%, such as at least 97%, such as at least98%, such as at least 99% or 100% sequence identity hereto. In oneaspect, the hexosaminidase is a polypeptide with SEQ ID NO 13 orpolypeptides having at least 60%, such as at least 65%, such as at least70%, such as at least 75%, such as at least 80%, such as at least 85%,or such as at least 90%, such as at least 91%, such as at least 92%,such as at least 93%, such as at least 94%, such as at least 95%, suchas at least 96%, such as at least 97%, such as at least 98%, such as atleast 99% or 100% sequence identity hereto. In one aspect, thehexosaminidase is a polypeptide with SEQ ID NO 14 or polypeptides havingat least 60%, such as at least 65%, such as at least 70%, such as atleast 75%, such as at least 80%, such as at least 85%, or such as atleast 90%, such as at least 91%, such as at least 92%, such as at least93%, such as at least 94%, such as at least 95%, such as at least 96%,such as at least 97%, such as at least 98%, such as at least 99% or 100%sequence identity hereto. In one aspect, the hexosaminidase is apolypeptide with SEQ ID NO 19 or polypeptides having at least 60%, suchas at least 65%, such as at least 70%, such as at least 75%, such as atleast 80%, such as at least 85%, or such as at least 90%, such as atleast 91%, such as at least 92%, such as at least 93%, such as at least94%, such as at least 95%, such as at least 96%, such as at least 97%,such as at least 98%, such as at least 99% or 100% sequence identityhereto. In one aspect, the hexosaminidase is a polypeptide with SEQ IDNO 20 or polypeptides having at least 60%, such as at least 65%, such asat least 70%, such as at least 75%, such as at least 80%, such as atleast 85%, or such as at least 90%, such as at least 91%, such as atleast 92%, such as at least 93%, such as at least 94%, such as at least95%, such as at least 96%, such as at least 97%, such as at least 98%,such as at least 99% or 100% sequence identity hereto.

Preferably the hexosaminidase is selected from the Curtobacterium cladeor the Terribacillus clade, most preferably the hexosaminidase isselected from the Terribacillus clade.

The enzymes having DNase activity to be used in the cleaning compositionof the invention, which are useful for deep cleaning of items such ashard surfaces, textiles and/or fabric, are preferably selected fromDNase which is obtainable from a fungus. A DNase which is obtainablefrom an Aspergillus is preferred; a DNase which is obtainable fromAspergillus oryzae is preferred. In one embodiment of the presentinvention, the enzyme having deoxyribonuclease activity is not the S1nuclease from Aspergillus oryzae.

The DNase used in the present invention preferably includes thepolypeptide having SEQ ID NO: 2, which is obtained from Aspergillusoryzae. One aspect of the present invention relates to isolated enzymeshaving a sequence identity to the mature polypeptide of SEQ ID NO: 2 ofat least 60%, e.g., at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100%, and which have DNase activity.

Biofilm can develop on textile when microorganisms are present on anitem and stick together on the item. This biofilm may adhere soil due toits sticky nature.

The present invention concerns the use of a cleaning compositioncomprising an enzyme having DNase activity and an enzyme havinghexosaminidase activity for deep cleaning of an item, wherein the DNaseis an enzyme having at least 60% sequence identity to the amino acidsequence having SEQ ID NO: 2, wherein the hexosaminidase is an enzymehaving at least 60% sequence identity to the amino acid sequence havingSEQ ID NO: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19or 20 and wherein the item is a textile. In one aspect of the inventionthe cleaning composition is used for preventing, reducing or removingthe stickiness of an item. The cleaning composition can further be usedfor pre-treating stains on textile such as textile with a pronouncedamount of biofilm adhered to the textile.

Additionally, the invention concerns use of the cleaning composition forpreventing, reducing or removing re-deposition of soil during a washcycle. When the cleaning composition is used for example in thelaundering of textile, the cleaning composition hinders deposition ofsoil present in the wash liquor on the textile.

Further, the invention concerns the use of a cleaning composition forpreventing, reducing or removing the adherence of soil to an item. Inone embodiment, the item is textile. When the soil does not adhere tothe item, the item appears cleaner. Thus, the invention further concernsthe use of a cleaning composition according to the invention formaintaining or improving the whiteness of the item.

When items like T-shirts or sportswear are used, they are exposed tobacteria from the body of the user and from the rest of the environmentin which they are used. This may cause malodor on the item even afterthe item is washed. The present invention therefore also concernsremoval or reduction of malodor on textile. The malodor may be caused bybacteria producing compounds with an unpleasant smell. One example ofsuch unpleasant smelling compounds is E-2-nonenal. The malodor can bepresent on newly washed textile which is still wet, or the malodor canbe present on newly washed textile which has subsequently been dried.The malodor may also be present on textile which has been stored forsome time after wash. The present invention relates to reduction orremoval of malodor such as E-2-nonenal from wet or dry textile.

The cleaning composition according to the invention may comprise acleaning adjunct; the cleaning adjunct ingredient may be selected fromsurfactants and builders and/or chelators such as those described above.The adjunct ingredients may also be any of the following: a flocculatingaid, dye transfer inhibitors, enzymes, enzyme stabilizers, enzymeinhibitors, catalytic materials, bleach activators, hydrogen peroxide,sources of hydrogen peroxide, preformed peracids, polymeric dispersingagents, clay soil removal/anti-redeposition agents, brighteners, sudssuppressors, dyes, perfumes, structure elasticizing agents, fabricsofteners, carriers, hydrotropes, builders and co-builders, fabrichueing agents, anti-foaming agents, dispersants, processing aids, and/orpigments.

In one embodiment, the cleaning adjunct ingredient is a builder or aclay soil removal/anti-redeposition agent.

In one embodiment, the cleaning adjunct ingredient is one or moreenzymes. The one or more enzymes may be selected from the groupconsisting of proteases, lipases, cutinases, amylases, carbohydrases,cellulases, pectinases, mannanases, arabinases, galactanases, xylanasesand oxidases.

In addition to the enzymes having DNase activity and the enzyme havinghexosaminidase activity the detergents of the invention may furthercomprise cellulases. Suitable cellulases include those of bacterial orfungal origin. Chemically modified or protein engineered mutants areincluded. Suitable cellulases include cellulases from the generaBacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g.,the fungal cellulases produced from Humicola insolens, Myceliophthorathermophila and Fusarium oxysporum disclosed in U.S. Pat. Nos.4,435,307, 5,648,263, 5,691,178, 5,776,757 and WO 89/09259.

Especially suitable cellulases are the alkaline or neutral cellulaseshaving color care benefits. Examples of such cellulases are cellulasesdescribed in EP 0 495 257, EP 0 531 372, WO 96/11262, WO 96/29397, WO98/08940. Other examples are cellulase polypeptides such as thosedescribed in WO 94/07998, EP 0 531 315, U.S. Pat. Nos. 5,457,046,5,686,593, 5,763,254, WO 95/24471, WO 98/12307 and PCT/DK98/00299.

Examples of cellulases exhibiting endo-beta-1,4-glucanase activity (EC3.2.1.4) are those described in WO02/099091.

Other examples of cellulases include the family 45 cellulases describedin WO96/29397, and especially polypeptides thereof having substitution,insertion and/or deletion at one or more of the positions correspondingto the following positions in SEQ ID NO: 8 of WO 02/099091: 2, 4, 7, 8,10, 13, 15, 19, 20, 21, 25, 26, 29, 32, 33, 34, 35, 37, 40, 42, 42a, 43,44, 48, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 70, 72, 76, 79, 80, 82,84, 86, 88, 90, 91, 93, 95, 95d, 95h, 95j, 97, 100, 101, 102, 103, 113,114, 117, 119, 121, 133, 136, 137, 138, 139, 140a, 141, 143a, 145, 146,147, 150e, 150j, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160c,160e, 160k, 161, 162, 164, 165, 168, 170, 171, 172, 173, 175, 176, 178,181, 183, 184, 185, 186, 188, 191, 192, 195, 196, 200, and/or 20,preferably selected among P19A, G20K, Q44K, N48E, Q119H or Q146R.

Commercially available cellulases include Celluzyme™, Celluclean andCarezyme™ (Novozymes NS), Clazinase™, and Puradax HA™ (GenencorInternational Inc.), and KAC-500(B)™ (Kao Corporation).

In addition to the enzymes having DNase activity and the enzyme havinghexosaminidase activity the cleaning composition of the invention mayfurther comprise proteases. Suitable proteases include those ofbacterial, fungal, plant, viral or animal origin e.g. vegetable ormicrobial origin. Microbial origin is preferred. Chemically modified orprotein engineered mutants are included. It may be an alkaline protease,such as a serine protease or a metalloprotease. A serine protease mayfor example be of the S1 family, such as trypsin, or the S8 family suchas subtilisin. A metalloprotease may for example be a thermolysin frome.g. family M4 or other metalloprotease such as those from the M5, M7 orM8 families.

The term “subtilases” refers to a sub-group of serine protease accordingto Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al.Protein Science 6 (1997) 501-523. Serine proteases are a subgroup ofproteases characterized by having a serine in the active site, whichforms a covalent adduct with the substrate. The subtilases may bedivided into 6 sub-divisions, i.e. the Subtilisin family, the Thermitasefamily, the Proteinase K family, the Lantibiotic peptidase family, theKexin family and the Pyrolysin family.

Examples of subtilases are those derived from Bacillus such as Bacilluslentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, Bacilluspumilus and Bacillus gibsonii described in U.S. Pat. No. 7,262,042 andWO09/021867, and subtilisin lentus, subtilisin Novo, subtilisinCarlsberg, Bacillus licheniformis, subtilisin BPN′, subtilisin 309,subtilisin 147 and subtilisin 168 described in WO89/06279 and proteasePD138 described in (WO93/18140). Other useful proteases may be thosedescribed in WO92/175177, WO01/016285, WO02/026024 and WO02/016547.Examples of trypsin-like proteases are trypsin (e.g. of porcine orbovine origin) and the Fusarium protease described in WO89/06270,WO94/25583 and WO05/040372, and the chymotrypsin proteases derived fromCellumonas described in WO05/052161 and WO05/052146.

A further preferred protease is the alkaline protease from Bacilluslentus DSM 5483, as described for example in WO95/23221, and variantsthereof which are described in WO92/21760, WO95/23221, EP1921147 andEP1921148.

Examples of metalloproteases are described in WO07/044993 (GenencorInt.) such as those derived from Bacillus amyloliquefaciens.

Examples of useful proteases are the variants described in: WO92/19729,WO96/034946, WO98/20115, WO98/20116, WO99/011768, WO01/44452,WO03/006602, WO04/03186, WO04/041979, WO07/006305, WO11/036263,WO11/036264, especially the variants with substitutions in one or moreof the following positions: 3, 4, 9, 15, 24, 27, 42, 55, 59, 60, 66, 74,85, 96, 97, 98, 99, 100, 101, 102, 104, 116, 118, 121, 126, 127, 128,154, 156, 157, 158, 161, 164, 176, 179, 182, 185, 188, 189, 193, 198,199, 200, 203, 206, 211, 212, 216, 218, 226, 229, 230, 239, 246, 255,256, 268 and 269, wherein the positions correspond to the positions ofthe Bacillus lentus protease shown in SEQ ID NO 1 of WO 2016/001449.More preferred the subtilase variants may comprise one or more of themutations: S3T, V4I, S9R, S9E, A15T, S24G, S24R, K27R, N42R, S55P, G59E,G59D, N60D, N60E, V66A, N74D, N85S, N85R, G96S, G96A, S97G, S97D, S97A,S97SD, S99E, S99D, S99G, S99M, S99N, S99R, S99H, S101A, V1021, V102Y,V102N, S104A, G116V, G116R, H118D, H118N, N1205, S126L, P127Q, S128A,S154D, A156E, G157D, G157P, S158E, Y161A, R164S, Q176E, N179E, S182E,Q185N, A188P, G189E, V193M, N198D, V199I, Y203W, S206G, L211Q, L211D,N212D, N212S, M216S, A226V, K229L, Q230H, Q239R, N246K, N255W, N255D,N255E, L256E, L256D T268A, R269H, wherein the positions correspond tothe positions of the Bacillus lentus protease shown in SEQ ID NO 1 of WO2016/001449. The protease variants are preferably variants of theBacillus lentus protease (Savinase®) shown in SEQ ID NO 1 of WO2016/001449, the Bacillus amylolichenifaciens protease (BPN′) shown inSEQ ID NO 2 of WO2016/001449. The protease variants preferably have atleast 80% sequence identity to SEQ ID NO 1 or SEQ ID NO 2 of WO2016/001449.

Another protease variant is one comprising a substitution at one or morepositions corresponding to positions 171, 173, 175, 179, or 180 of SEQID NO: 1 of WO2004/067737, wherein said protease variant has a sequenceidentity of at least 75% but less than 100% to SEQ ID NO: 1 ofWO2004/067737.

Suitable commercially available protease enzymes include those soldunder the trade names Alcalase®, Duralase™, Durazym™, Relase®, Relase®Ultra, Savinase®, Savinase® Ultra, Primase®, Polarzyme®, Kannase®,Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra,Blaze®, Blaze Evity® 100T, Blaze Evity® 125T, Blaze Evity® 150T,Neutrase®, Everlase® and Esperase® (Novozymes A/S), those sold under thetradename Maxatase®, Maxacal®, Maxapem®, Purafect Ox®, Purafect OxP®,Puramax®, FN2®, FN3®, FN4®, Excellase®, Excellenz P1000™, ExcellenzP1250™, Eraser®, Preferenz P100™, Purafect Prime®, Preferenz P110™,Effectenz P1000™, Purafect®™, Effectenz P1050™, Purafect Ox®™, EffectenzP2000™, Purafast®, Properase®, Opticlean® and Optimase®(Danisco/DuPont), Axapem™ (Gist-Brocases N.V.), BLAP (sequence shown inFIG. 29 of U.S. Pat. No. 5,352,604) and variants hereof (Henkel AG) andKAP (Bacillus alkalophilus subtilisin) from Kao.

In addition to the enzymes having DNase activity and the enzyme havinghexosaminidase activity the cleaning composition of the invention mayfurther comprise lipases and cutinases which include those of bacterialor fungal origin. Chemically modified or protein engineered mutantenzymes are included. Examples include lipase from Thermomyces, e.g.from T. lanuginosus (previously named Humicola lanuginosa) as describedin EP258068 and EP305216, cutinase from Humicola, e.g. H. insolens(WO96/13580), lipase from strains of Pseudomonas (some of these nowrenamed to Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes(EP218272), P. cepacia (EP331376), P. sp. strain SD705 (WO95/06720 &WO96/27002), P. wisconsinensis (WO96/12012), GDSL-type Streptomyceslipases (WO10/065455), cutinase from Magnaporthe grisea (WO10/107560),cutinase from Pseudomonas mendocina (U.S. Pat. No. 5,389,536), lipasefrom Thermobifida fusca (WO11/084412), Geobacillus stearothermophiluslipase (WO11/084417), lipase from Bacillus subtilis (WO11/084599), andlipase from Streptomyces griseus (WO11/150157) and S. pristinaespiralis(WO12/137147).

Other examples are lipase polypeptides such as those described inEP407225, WO92/05249, WO94/01541, WO94/25578, WO95/14783, WO95/30744,WO95/35381, WO95/22615, WO96/00292, WO97/04079, WO97/07202, WO00/34450,WO00/60063, WO01/92502, WO07/87508 and WO09/109500.

Preferred commercial lipase products include include Lipolase™, Lipex™;Lipolex™ and Lipoclean™ (Novozymes A/S), Lumafast (originally fromGenencor) and Lipomax (originally from Gist-Brocades).

Still other examples are lipases sometimes referred to asacyltransferases or perhydrolases, e.g. acyltransferases with homologyto Candida antarctica lipase A (WO10/111143), acyltransferase fromMycobacterium smegmatis (WO05/56782), perhydrolases from the CE 7 family(WO09/67279), and polypeptides of the M. smegmatis perhydrolase inparticular the S54V variant used in the commercial product Gentle PowerBleach from Huntsman Textile Effects Pte Ltd (WO10/100028).

In addition to the enzymes having DNase activity and the enzyme havinghexosaminidase activity the cleaning composition of the invention mayfurther comprise amylases. The amylase may be an alpha-amylase or aglucoamylase and may be of bacterial or fungal origin. Chemicallymodified or protein engineered mutants are included. Amylases include,for example, alpha-amylases obtained from Bacillus, e.g., a specialstrain of Bacillus licheniformis, described in more detail in GB1,296,839.

Suitable amylases include amylases having SEQ ID NO: 3 in WO 95/10603 orpolypeptides having 90% sequence identity to SEQ ID NO: 3 thereof.Preferred polypeptides are described in WO 94/02597, WO 94/18314, WO97/43424 and SEQ ID NO: 4 of WO 99/019467, such as polypeptides withsubstitutions in one or more of the following positions: 15, 23, 105,106, 124, 128, 133, 154, 156, 178, 179, 181, 188, 190, 197, 201, 202,207, 208, 209, 211, 243, 264, 304, 305, 391, 408, and 444.

Different suitable amylases include amylases having SEQ ID NO: 6 in WO02/010355 or polypeptides thereof having 90% sequence identity to SEQ IDNO: 6. Preferred polypeptides of SEQ ID NO: 6 are those having adeletion in positions 181 and 182 and a substitution in position 193.

Other amylases which are suitable are hybrid alpha-amylase comprisingresidues 1-33 of the alpha-amylase derived from B. amyloliquefaciensshown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of the B.licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO 2006/066594 orpolypeptides having 90% sequence identity thereof. Preferredpolypeptides of this hybrid alpha-amylase are those having asubstitution, a deletion or an insertion in one of more of the followingpositions: G48, T49, G107, H156, A181, N190, M197, I201, A209 and Q264.Most preferred polypeptides of the hybrid alpha-amylase comprisingresidues 1-33 of the alpha-amylase derived from B. amyloliquefaciensshown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of SEQ IDNO: 4 are those having the substitutions:

M197T;

H156Y+A181T+N190F+A209V+Q264S; or

G48A+T49I+G107A+H156Y+A181T+N190F+1201F+A209V+Q264S.

Further amylases which are suitable are amylases having SEQ ID NO: 6 inWO 99/019467 or polypeptides thereof having 90% sequence identity to SEQID NO: 6. Preferred polypeptides of SEQ ID NO: 6 are those having asubstitution, a deletion or an insertion in one or more of the followingpositions: R181, G182, H183, G184, N195, I206, E212, E216 and K269.Particularly preferred amylases are those having deletion in positionsR181 and G182, or positions H183 and G184.

Additional amylases which can be used are those having SEQ ID NO: 1, SEQID NO: 3, SEQ ID NO: 2 or SEQ ID NO: 7 of WO 96/023873 or polypeptidesthereof having 90% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQID NO: 3 or SEQ ID NO: 7. Preferred polypeptides of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3 or SEQ ID NO: 7 are those having a substitution, adeletion or an insertion in one or more of the following positions: 140,181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304 and 476. Morepreferred polypeptides are those having a deletion in positions 181 and182 or positions 183 and 184. Most preferred amylase polypeptides of SEQID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7 are those having a deletion inpositions 183 and 184 and a substitution in one or more of positions140, 195, 206, 243, 260, 304 and 476.

Other amylases which can be used are amylases having SEQ ID NO: 2 of WO08/153815, SEQ ID NO: 10 in WO 01/66712 or polypeptides thereof having90% sequence identity to SEQ ID NO: 2 of WO 08/153815 or 90% sequenceidentity to SEQ ID NO: 10 in WO 01/66712. Preferred polypeptides of SEQID NO: 10 in WO 01/66712 are those having a substitution, a deletion oran insertion in one of more of the following positions: 176, 177, 178,179, 190, 201, 207, 211 and 264.

Further suitable amylases are amylases having SEQ ID NO: 2 of WO09/061380 or polypeptides having 90% sequence identity to SEQ ID NO: 2thereof. Preferred polypeptides of SEQ ID NO: 2 are those having atruncation of the C-terminus and/or a substitution, a deletion or aninsertion in one of more of the following positions: Q87, Q98, S125,N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243,N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferredpolypeptides of SEQ ID NO: 2 are those having the substitution in one ofmore of the following positions: Q87E,R, Q98R, S125A, N128C, T131I,T165I, K178L, T182G, M201L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R,R309A, Q320R, Q359E, K444E and G475K and/or deletion in position R180and/or S181 or of T182 and/or G183. Most preferred amylase polypeptidesof SEQ ID NO: 2 are those having the substitutions:

N128C+K178L+T182G+Y305R+G475K;

N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;

S125A+N128C+K178L+T182G+Y305R+G475K; or

S125A+N128C+T131I+T165I+K178L+T182G+Y305R+G475K, wherein thepolypeptides are C-terminally truncated and optionally further comprisea substitution at position 243 and/or a deletion at position 180 and/orposition 181.

Other suitable amylases are the alpha-amylase having SEQ ID NO: 12 inWO01/66712 or a variant having at least 90% sequence identity to SEQ IDNO: 12. Preferred amylase polypeptides are those having a substitution,a deletion or an insertion in one of more of the following positions ofSEQ ID NO: 12 in WO01/66712: R28, R118, N174; R181, G182, D183, G184,G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320,H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484.Particular preferred amylases include polypeptides having a deletion ofD183 and G184 and having the substitutions R118K, N195F, R320K andR458K, and a variant additionally having substitutions in one or moreposition selected from the group: M9, G149, G182, G186, M202, T257,Y295, N299, M323, E345 and A339, most preferred a variant thatadditionally has substitutions in all these positions.

Other examples are amylase polypeptides such as those described inWO2011/098531, WO2013/001078 and WO2013/001087.

Commercially available amylases are Duramyl™, Termamyl™, Fungamyl™,Stainzyme™, Stainzyme Plus™, Natalase™, Liquozyme X and BAN™ (fromNovozymes NS), and Rapidase™, Purastar™/Effectenz™, Powerase andPreferenz S100 (from Genencor International Inc./DuPont).

In addition to the enzymes having DNase activity and the enzyme havinghexosaminidase activity the cleaning composition of the invention mayfurther comprise peroxidases/oxidases including those of plant,bacterial or fungal origin. Chemically modified or protein engineeredmutants are included. Examples of useful peroxidases include peroxidasesfrom Coprinus, e.g., from C. cinereus, and variants thereof as thosedescribed in WO 93/24618, WO 95/10602, and WO 98/15257.

Commercially available peroxidases include Guardzyme™ (Novozymes A/S).

The detergent or cleaning composition enzyme(s) may be included in adetergent composition by adding separate additives containing one ormore enzymes, or by adding a combined additive comprising these enzymes.A detergent additive of the invention, i.e., a separate additive or acombined additive, can be formulated, for example, as a granulate,liquid, slurry, etc. Preferred detergent additive formulations aregranulating, non-dusting granulates, liquids, stabilized liquids, orslurries.

Non-dusting granulates may be produced, e.g., as disclosed in U.S. Pat.Nos. 4,106,991 and 4,661,452 and may optionally be coated by methodsknown in the art. Examples of waxy coating materials are poly (ethyleneoxide) products (polyethyleneglycol, PEG) with mean molar weights of1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethyleneoxide units; ethoxylated fatty alcohols in which the alcohol containsfrom 12 to 20 carbon atoms and in which there are 15 to 80 ethyleneoxide units; fatty alcohols; fatty acids; and mono- and di- andtriglycerides of fatty acids. Examples of film-forming coating materialssuitable for application by fluid bed techniques are given in GB1483591. Liquid enzyme preparations may, for instance, be stabilized byadding a polyol such as propylene glycol, a sugar or sugar alcohol,lactic acid or boric acid according to established methods. Protectedenzymes may be prepared according to the method disclosed in EP 238,216.

The cleaning compositions of the invention may also contain 0-10% byweight, such as 0.5-5%, 2-5%, 0.5-2% or 0.2-1% of a polymer. Any polymerknown in the art for use in detergents may be utilized. The polymer mayfunction as a co-builder as mentioned above, or may provideantiredeposition, fiber protection, soil release, dye transferinhibition, grease cleaning and/or anti-foaming properties. Somepolymers may have more than one of the above-mentioned properties and/ormore than one of the below-mentioned motifs. Exemplary polymers include(carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA),poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethyleneoxide) (PEG), ethoxylated poly(ethyleneimine), (carboxymethyl)inulin(CMI), and polycarboxylates such as PAA, PAA/PMA, polyaspartic acid, andlauryl methacrylate/acrylic acid copolymers, hydrophobically modifiedCMC (HM-CMC) and silicones, copolymers of terephthalic acid andoligomeric glycols, copolymers of poly(ethylene terephthalate) andpoly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole)(PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) andpolyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymersinclude sulfonated polycarboxylates, polyethylene oxide andpolypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Otherexemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of theabove-mentioned polymers are also contemplated.

The cleaning compositions of the present invention may also includefabric hueing agents such as dyes or pigments, which when formulated indetergent compositions can deposit onto a fabric when said fabric iscontacted with a wash liquor comprising said detergent compositions andthus altering the tint of said fabric through absorption/reflection ofvisible light. Fluorescent whitening agents emit at least some visiblelight if subjected to ultraviolet light. In contrast, fabric hueingagents alter the tint of a surface as they absorb at least a portion ofthe visible light spectrum. Suitable fabric hueing agents include dyesand dye-clay conjugates, and may also include pigments. Suitable dyesinclude small molecule dyes and polymeric dyes. Suitable small moleculedyes include small molecule dyes selected from the group consisting ofdyes falling into the Colour Index (C.I.) classifications of DirectBlue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, BasicBlue, Basic Violet and Basic Red, or mixtures thereof, for example asdescribed in WO2005/03274, WO2005/03275, WO2005/03276 and EP1876226(hereby incorporated by reference). The cleaning composition preferablycomprises from about 0.00003 wt % to about 0.2 wt %, from about 0.00008wt % to about 0.05 wt %, or even from about 0.0001 wt % to about 0.04 wt% fabric hueing agent. The composition may comprise from 0.0001 wt % to0.2 wt % fabric hueing agent, this may be especially preferred when thecomposition is in the form of a unit dose pouch. Suitable hueing agentsare also disclosed in, e.g. WO 2007/087257 and WO2007/087243.

The cleaning may contain 0-10% by weight, for example 0-5% by weight,such as about 0.5 to about 5%, or about 3% to about 5%, of a hydrotrope.Any hydrotrope known in the art for use in detergents may be utilized.Non-limiting examples of hydrotropes include sodium benzenesulfonate,sodium p-toluene sulfonate (STS), sodium xylene sulfonate (SXS), sodiumcumene sulfonate (SCS), sodium cymene sulfonate, amine oxides, alcoholsand polyglycolethers, sodium hydroxynaphthoate, sodiumhydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, andcombinations thereof.

The cleaning compositions of the present invention can also containdispersants. In particular, powdered detergents may comprisedispersants. Suitable water-soluble organic materials include the homo-or co-polymeric acids or their salts, in which the polycarboxylic acidcomprises at least two carboxyl radicals separated from each other bynot more than two carbon atoms. Suitable dispersants are for exampledescribed in Powdered Detergents, Surfactant science series volume 71,Marcel Dekker, Inc.

The cleaning compositions of the present invention may also include oneor more dye transfer inhibiting agents. Suitable polymeric dye transferinhibiting agents include, but are not limited to, polyvinylpyrrolidonepolymers, polyamine-N-oxide polymers, copolymers of N-vinylpyrrolidoneand N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles ormixtures thereof. When present in a subject composition, the dyetransfer inhibiting agents may be present at levels from about 0.0001%to about 10%, from about 0.01% to about 5% or even from about 0.1% toabout 3% by weight of the composition.

The cleaning compositions of the present invention will preferably alsocontain additional components that may tint articles being cleaned, suchas fluorescent whitening agents or optical brighteners. Where presentthe brightener is preferably at a level of about 0.01% to about 0.5% byweight. Any fluorescent whitening agent suitable for use in a laundrydetergent composition may be used in the cleaning composition of thepresent invention. The most commonly used fluorescent whitening agentsare those belonging to the classes of diaminostilbene-sulfonic acidderivatives, diarylpyrazoline derivatives and biphenyl-distyrylderivatives. Examples of the diaminostilbene-sulfonic acid derivativetype of fluorescent whitening agents include the sodium salts of:4,4′-bis[(4-anilino-6-diethanolamino-s-triazin-2-yl)amino]stilbene-2,2′-disulfonate,4,4′-bis[(4,6-dianilino-s-triazin-2-yl)amino]stilbene-2,2′-disulfonate,4,4′-bis{4-anilino-6-[methyl(2-hydroxyethyl)amino]-s-triazin-2-ylamino}stilbene-2,2′-disulfonate,4,4′-bis(4-phenyl-1,2,3-triazol-2-yl)stilbene-2,2′-disulfonate andsodium5-(2H-naphtho[1,2-d][1,2,3]triazol-2-yl)-2-[(E)-2-phenylvinyl]benzenesulfonate.Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBSavailable from BASF. Tinopal DMS is the disodium salt of4,4′-bis[(4-anilino-6-morpholino-s-triazin-2-yl) amino]stilbene-2,2′-disulfonate. Tinopal CBS is the disodium salt of2,2′-[biphenyls-4,4′-di(2,1-ethenediyl)] dibenzene-1-sulfonate. Alsopreferred is the commercially available Parawhite KX, supplied byParamount Minerals and Chemicals, Mumbai, India. Other fluorescerssuitable for use include the 1-3-diarylpyrazolines and the7-alkylaminocoumarins.

Suitable fluorescent brightener levels include lower levels of fromabout 0.01, from 0.05, from about 0.1 or even from about 0.2 wt % toupper levels of 0.5 or even 0.75 wt %.

The cleaning compositions of the present invention may also include oneor more soil-release polymers which aid the removal of soils fromfabrics such as cotton and polyester-based fabrics, the removal ofhydrophobic soils from polyester-based fabrics. The soil releasepolymers may for example be nonionic or anionic terephthalate-basedpolymers, polyvinylcaprolactam and related copolymers, vinyl graftcopolymers or polyester polyamides; see for example Chapter 7 inPowdered Detergents, Surfactant science series volume 71, Marcel Dekker,Inc. Another type of soil release polymers is amphiphilic alkoxylatedgrease cleaning polymers comprising a core structure and a plurality ofalkoxylate groups attached to that core structure. The core structuremay comprise a polyalkylenimine structure or a polyalkanolaminestructure as described in detail in WO 2009/087523 (hereby incorporatedby reference). Furthermore, random graft co-polymers are suitablesoil-release polymers. Suitable graft co-polymers are described in moredetail in WO 2007/138054, WO 2006/108856 and WO 2006/113314 (herebyincorporated by reference). Other soil-release polymers are substitutedpolysaccharide structures especially substituted cellulosic structuressuch as modified cellulose derivatives such as those described in EP1867808 or WO 2003/040279 (both are hereby incorporated by reference).Suitable cellulosic polymers include cellulose, cellulose ethers,cellulose esters, cellulose amides and mixtures thereof. Suitablecellulosic polymers include anionically modified cellulose, nonionicallymodified cellulose, cationically modified cellulose, zwitterionicallymodified cellulose, and mixtures thereof.

The cleaning compositions of the present invention may also include oneor more anti-redeposition agents such as (carboxymethyl) cellulose(CMC), poly (vinyl alcohol) (PVA), homopolymers of acrylic acid,copolymers of acrylic acid and maleic acid, and ethoxylatedpolyethyleneimines. The cellulose based polymers described undersoil-release polymers above may also function as anti-redepositionagents.

The cleaning composition of the invention may also contain one are moreadjunct materials. Suitable adjunct materials include, but are notlimited to, anti-shrink agents, anti-wrinkling agents, bactericides,binders, carriers, dyes, enzyme stabilizers, fabric softeners, fillers,foam regulators, hydrotropes, perfumes, pigments, sod suppressors,solvents, and structurants for liquid detergents and/or structureelasticizing agents.

Formulation of Cleaning Products

The cleaning composition of the invention, e.g. a detergent composition,may be in any convenient form, e.g., a bar, a homogenous tablet, atablet having two or more layers, a pouch having one or morecompartments, a regular or compact powder, a granule, a paste, a gel, ora regular, compact or concentrated liquid.

Pouches can be configured as single or multicompartments. It can be ofany form, shape and material which is suitable for hold the composition,e.g. without allowing the release of the composition to release of thecomposition from the pouch prior to water contact. The pouch is madefrom water soluble film which encloses an inner volume. Said innervolume can be divided into compartments of the pouch. Preferred filmsare polymeric materials preferably polymers which are formed into a filmor sheet. Preferred polymers, copolymers or derivates thereof areselected polyacrylates, and water soluble acrylate copolymers, methylcellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose,hydroxyethyl cellulose, hydroxypropyl methyl cellulose, malto dextrin,poly methacrylates, most preferably polyvinyl alcohol copolymers and,hydroxypropyl methyl cellulose (HPMC). Preferably the level of polymerin the film for example PVA is at least about 60%. Preferred averagemolecular weight will typically be about 20,000 to about 150,000. Filmscan also be of blended compositions comprising hydrolytically degradableand water soluble polymer blends such as polylactide and polyvinylalcohol (known under the Trade reference M8630 as sold by MonoSol LLC,Indiana, USA) plus plasticisers like glycerol, ethylene glycerol,propylene glycol, sorbitol and mixtures thereof. The pouches cancomprise a solid laundry cleaning composition or part components and/ora liquid cleaning composition or part components separated by thewater-soluble film. The compartment for liquid components can bedifferent in composition than compartments containing solids:US2009/0011970 A1.

Cleaning ingredients, such as detergent ingredients, can be separatedphysically from each other by compartments in water dissolvable pouchesor in different layers of tablets. Thereby negative storage interactionbetween components can be avoided. Different dissolution profiles ofeach of the compartments can also give rise to delayed dissolution ofselected components in the wash solution.

A liquid or gel detergent which is not unit dosed may be aqueous,typically containing at least 20% by weight and up to 95% water, such asup to about 70% water, up to about 65% water, up to about 55% water, upto about 45% water, up to about 35% water. Other types of liquids,including without limitation, alkanols, amines, diols, ethers andpolyols may be included in an aqueous liquid or gel. An aqueous liquidor gel detergent may contain from 0-30% organic solvent.

A liquid or gel detergent may be non-aqueous.

Laundry Soap Bars

The enzyme having DNase activity and the enzyme having hexosaminidaseactivity may be added to laundry soap bars and used for hand washinglaundry, fabrics and/or textiles. The term laundry soap bar includeslaundry bars, soap bars, combo bars, syndet bars and detergent bars. Thetypes of bar usually differ in the type of surfactant they contain, andthe term laundry soap bar includes those containing soaps from fattyacids and/or synthetic soaps. The laundry soap bar has a physical formwhich is solid and not a liquid, gel or a powder at room temperature.The term solid is defined as a physical form which does notsignificantly change over time, i.e. if a solid object (e.g. laundrysoap bar) is placed inside a container, the solid object does not changeto fill the container it is placed in. The bar is a solid typically inbar form but can be in other solid shapes such as round or oval.

The laundry soap bar may contain one or more additional enzymes,protease inhibitors such as peptide aldehydes (or hydrosulfite adduct orhemiacetal adduct), boric acid, borate, borax and/or phenylboronic acidderivatives such as 4-formylphenylboronic acid, one or more soaps orsynthetic surfactants, polyols such as glycerine, pH controllingcompounds such as fatty acids, citric acid, acetic acid and/or formicacid, and/or a salt of a monovalent cation and an organic anion whereinthe monovalent cation may be for example Na⁺, K⁺ or NH₄ ⁺ and theorganic anion may be for example formate, acetate, citrate or lactatesuch that the salt of a monovalent cation and an organic anion may be,for example, sodium formate.

The laundry soap bar may also contain complexing agents like EDTA andHEDP, perfumes and/or different type of fillers, surfactants e.g.anionic synthetic surfactants, builders, polymeric soil release agents,detergent chelators, stabilizing agents, fillers, dyes, colorants, dyetransfer inhibitors, alkoxylated polycarbonates, suds suppressers,structurants, binders, leaching agents, bleaching activators, clay soilremoval agents, anti-redeposition agents, polymeric dispersing agents,brighteners, fabric softeners, perfumes and/or other compounds known inthe art.

The laundry soap bar may be processed in conventional laundry soap barmaking equipment such as but not limited to: mixers, plodders, e.g a twostage vacuum plodder, extruders, cutters, logo-stampers, cooling tunnelsand wrappers. The invention is not limited to preparing the laundry soapbars by any single method. The premix may be added to the soap atdifferent stages of the process. For example, the premix containing asoap, enzymes of the invention, optionally one or more additionalenzymes, a protease inhibitor, and a salt of a monovalent cation and anorganic anion may be prepared and the mixture is then plodded. Theenzymes of the invention and optional additional enzymes may be added atthe same time as the protease inhibitor for example in liquid form.Besides the mixing step and the plodding step, the process may furthercomprise the steps of milling, extruding, cutting, stamping, coolingand/or wrapping.

Formulation of Enzyme in Co-Granule

The enzyme having DNase activity and the enzyme having hexosaminidaseactivity may be formulated as a granule for example as a co-granule thatcombines one or more enzymes. Each enzyme will then be present in moregranules securing a more uniform distribution of enzymes in thedetergent. This also reduces the physical segregation of differentenzymes due to different particle sizes. Methods for producingmulti-enzyme co-granulate for the detergent industry are disclosed inthe IP.com disclosure IPCOM000200739D.

Another example of formulation of enzymes using co-granulates isdisclosed in WO 2013/188331, which relates to a detergent compositioncomprising (a) a multi-enzyme co-granule; (b) less than 10 wt zeolite(anhydrous basis); and (c) less than 10 wt phosphate salt (anhydrousbasis), wherein said enzyme co-granule comprises from 10 to 98 wt %moisture sink components and the composition additionally comprises from20 to 80 wt % detergent moisture sink components. WO 2013/188331 alsorelates to a method of treating and/or cleaning a surface, preferably afabric surface comprising the steps of (i) contacting said surface withthe detergent composition as claimed and described herein in aqueouswash liquor, (ii) rinsing and/or drying the surface.

The multi-enzyme co-granule may comprise a enzymes of the invention and(a) one or more enzymes selected from the group consisting of first-washlipases, cleaning cellulases, xyloglucanases, perhydrolases,peroxidases, lipoxygenases, laccases and mixtures thereof; and (b) oneor more enzymes selected from the group consisting of hemicellulases,proteases, care cellulases, cellobiose dehydrogenases, xylanases,phospho lipases, esterases, cutinases, pectinases, mannanases, pectatelyases, keratinases, reductases, oxidases, phenoloxidases, ligninases,pullulanases, tannases, pentosanases, lichenases glucanases,arabinosidases, hyaluronidase, chondroitinase, amylases, and mixturesthereof.

The invention is further summarized in the following paragraphs:

-   1. Use of a combination of an enzyme having DNase activity and an    enzyme having hexosaminidase activity for deep cleaning of an item,    wherein the item is a textile.-   2. Use according to paragraph 1 for preventing, reducing or removing    stickiness of the item.-   3. Use according to any of paragraphs 1 or 2 for pre-treating stains    on the item.-   4. Use according to any of paragraphs 1-3 for preventing, reducing    or removing re-deposition of soil during a wash cycle.-   5. Use according to any of paragraphs 1-4 for preventing, reducing    or removing adherence of soil to the item.-   6. Use according to any of the preceding paragraphs for maintaining    or improving the whiteness of the item.-   7. Use according to any of the preceding paragraphs, wherein a    malodor is reduced or removed from the item.-   8. Use according to any of the preceding paragraphs, wherein the    textile is made of cotton, Cotton/Polyester, Polyester, Polyamide,    Polyacryl and/or silk.-   9. Use of a combination of an enzyme having DNase activity and an    enzyme having hexosaminidase activity for deep cleaning of an item,    wherein the item is a hard surface.-   10. Use according to any of the preceding paragraphs, wherein the    enzyme having hexosaminidase activity is a dispersin.-   11. Use according to any of the preceding paragraphs, wherein the    enzyme having hexosaminidase activity is selected from the group    consisting of DspB clade, the Curtobacterium clade and the    Terribacillus clade.-   12. Use according to any of paragraphs 10 or 11, wherein the enzyme    having hexosaminidase activity is selected from the DspB clade.-   13. Use according to paragraph 12, wherein the enzyme having    hexosaminidase activity is selected from polypeptides having at    least 60%, such as at least 65%, such as at least 70%, such as at    least 75%, such as at least 80%, such as at least 85%, or such as at    least 90%, such as at least 91%, such as at least 92%, such as at    least 93%, such as at least 94%, such as at least 95%, such as at    least 96%, such as at least 97%, such as at least 98%, such as at    least 99% or 100% sequence identity the amino acid sequences with    SEQ ID NOS 3, 4, 5, 6, 7, 8, 9, 10.-   14. Use according to any of paragraphs 10 or 11, wherein the enzyme    having hexosaminidase activity is selected from the Curtobacterium    clade.-   15. Use according to paragraph 14, wherein the enzyme having    hexosaminidase activity is a polypeptide having at least 60%, such    as at least 65%, such as at least 70%, such as at least 75%, such as    at least 80%, such as at least 85%, or such as at least 90%, such as    at least 91%, such as at least 92%, such as at least 93%, such as at    least 94%, such as at least 95%, such as at least 96%, such as at    least 97%, such as at least 98%, such as at least 99% or 100%    sequence identity to the amino acid sequence with SEQ ID NO 11, 15,    16, 17 or 18.-   16. Use according to any of paragraphs 10 or 11, wherein the enzyme    having hexosaminidase activity is selected from the Terribacillus    clade.-   17. Use according to paragraph 16, wherein the enzyme having    hexosaminidase activity is selected from polypeptides having at    least 60%, such as at least 65%, such as at least 70%, such as at    least 75%, such as at least 80%, such as at least 85%, or such as at    least 90%, such as at least 91%, such as at least 92%, such as at    least 93%, such as at least 94%, such as at least 95%, such as at    least 96%, such as at least 97%, such as at least 98%, such as at    least 99% or 100% sequence identity the amino acid sequences with    SEQ ID NO 12, 13, 14, 19 or 20.-   18. Use according to any of the preceding claims, wherein the enzyme    having DNase activity is a fungal DNase.-   19. Use according to paragraph 18, wherein the enzyme having DNase    activity is obtained from Aspergillus.-   20. Use according to paragraph 19, wherein the enzyme having DNase    activity is obtained from Aspergillus oryzae.-   21. Use according to any of the paragraphs 18 to 20, wherein the    enzyme having DNase activity is a polypeptide having a sequence    identity to the polypeptide of SEQ ID NO: 2 of at least 60%, e.g.,    at least 65%, at least 70%, at least 75%, at least 80%, at least    85%, at least 90%, at least 91%, at least 92%, at least 93%, at    least 94%, at least 95%, at least 96%, at least 97%, at least 98%,    at least 99%, or 100%.-   22. A composition comprising an enzyme having DNase activity and an    enzyme having hexosaminidase activity and an adjunct ingredient,    such as a cleaning agent.-   23. The composition according to paragraph 22, wherein the adjunct    ingredient is selected from the group consisting of surfactants,    builders, flocculating aid, chelating agents, dye transfer    inhibitors, enzymes, enzyme stabilizers, enzyme inhibitors,    catalytic materials, bleach activators, hydrogen peroxide, sources    of hydrogen peroxide, preformed peracids, polymeric dispersing    agents, clay soil removal/anti-redeposition agents, brighteners,    suds suppressors, dyes, perfumes, structure elasticizing agents,    fabric softeners, carriers, hydrotropes, builders and co-builders,    fabric huing agents, anti-foaming agents, dispersants, processing    aids, and/or pigments.-   24. The composition according to any of the preceding composition    paragraphs wherein the composition comprises from about 2 wt % to    about 50 wt %, 5 wt % to about 50 wt %, from about 5 wt % to about    40 wt %, from about 5 wt % to about 30 wt %, from about 5 wt % to    about 20 wt %, from about 5 wt % to about 10 wt % anionic    surfactant, preferably selected from linear alkylbenzenesulfonates    (LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS),    phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin    sulfonates, alkene sulfonates, alkane-2,3-diylbis(sulfates),    hydroxyalkanesulfonates and disulfonates, alkyl sulfates (AS) such    as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS),    primary alcohol sulfates (PAS), alcohol ethersulfates (AES or AEOS    or FES), secondary alkanesulfonates (SAS), paraffin sulfonates (PS),    ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo    fatty acid methyl esters (alpha-SFMe or SES) including methyl ester    sulfonate (MES), alkyl- or alkenylsuccinic acid,    dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives    of amino acids, diesters and monoesters of sulfo-succinic acid or    salt of fatty acids (soap), and combinations thereof.-   25. The composition according to any of the preceding composition    paragraphs wherein the composition comprises from about 2 wt % to    about 50 wt %, 5 wt % to about 50 wt %, 10 wt % to about 50 wt % of    at least one builder, preferably selected from citric acid,    methylglycine-N, N-diacetic acid (MGDA) and/or glutamic acid-N,    N-diacetic acid (GLDA) and mixtures thereof.-   26. The composition according to any of the preceding composition    paragraphs wherein the composition comprises at least one bleach    component, preferably a percarbonate and a manganese catalyst,    preferably 1,4,7-trimethyl-1,4,7-triazacyclononane or    manganese (III) acetate tetrahydrate (MnTACN).-   27. The composition according to any of the preceding paragraphs,    wherein the enzyme having hexosaminidase activity is a dispersin.-   28. The composition according to any of the preceding paragraphs,    wherein the enzyme having hexosaminidase activity is selected from    the group consisting of DspB clade, the Curtobacterium clade and the    Terribacillus clade.-   29. The composition according to any of paragraphs 27 or 28, wherein    the enzyme having hexosaminidase activity is selected from the DspB    clade.-   30. The composition according to paragraph 29, wherein the enzyme    having hexosaminidase activity is selected from polypeptides having    at least 60%, such as at least 65%, such as at least 70%, such as at    least 75%, such as at least 80%, such as at least 85%, or such as at    least 90%, such as at least 91%, such as at least 92%, such as at    least 93%, such as at least 94%, such as at least 95%, such as at    least 96%, such as at least 97%, such as at least 98%, such as at    least 99% or 100% sequence identity the amino acid sequences with    SEQ ID NOS 3, 4, 5, 6, 7, 8, 9 and 10.-   31. The composition according to any of paragraphs 27 or 28, wherein    the enzyme having hexosaminidase activity is selected from the    Curtobacterium clade.-   32. The composition according to paragraph 31, wherein the enzyme    having hexosaminidase activity is a polypeptide having at least 60%,    such as at least 65%, such as at least 70%, such as at least 75%,    such as at least 80%, such as at least 85%, or such as at least 90%,    such as at least 91%, such as at least 92%, such as at least 93%,    such as at least 94%, such as at least 95%, such as at least 96%,    such as at least 97%, such as at least 98%, such as at least 99% or    100% sequence identity to the amino acid sequence with SEQ ID NO 11,    15, 16, 17 or 18.-   33. The composition according to any of paragraphs 27 or 28, wherein    the enzyme having hexosaminidase activity is selected from the    Terribacillus clade.-   34. The composition according to paragraph 33, wherein the enzyme    having hexosaminidase activity is selected from polypeptides having    at least 60%, such as at least 65%, such as at least 70%, such as at    least 75%, such as at least 80%, such as at least 85%, or such as at    least 90%, such as at least 91%, such as at least 92%, such as at    least 93%, such as at least 94%, such as at least 95%, such as at    least 96%, such as at least 97%, such as at least 98%, such as at    least 99% or 100% sequence identity to the amino acid sequences with    SEQ ID NOS 12, 13, 14, 19 or 20.-   35. The composition according to any of the preceding claims,    wherein the enzyme having DNase activity is a fungal DNase.-   36. The composition according to paragraph 35, wherein the enzyme    having DNase activity is obtained from Aspergillus.-   37. The composition according to paragraph 36, wherein the enzyme    having DNase activity is obtained from Aspergillus oryzae.-   38. The composition according to any of the paragraphs 35 to 37,    wherein the enzyme having DNase activity is a polypeptide having a    sequence identity to the polypeptide of SEQ ID NO: 2 of at least    60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%,    at least 85%, at least 90%, at least 91%, at least 92%, at least    93%, at least 94%, at least 95%, at least 96%, at least 97%, at    least 98%, at least 99%, or 100%.-   39. The composition according to any of the preceding paragraphs    comprising from about 5 wt % to about 40 wt % nonionic surfactants,    and from about 0 wt % to about 50 wt % anionic surfactants.-   40. The composition according to paragraph 39, wherein the nonionic    surfactant is selected from alcohol ethoxylates (AE or AEO), alcohol    propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty    acid alkyl esters, such as ethoxylated and/or propoxylated fatty    acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol    ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines,    fatty acid monoethanolamides (FAM), fatty acid diethanolamides    (FADA), ethoxylated fatty acid monoethanolamides (EFAM),    propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl    fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine    (glucamides, GA, or fatty acid glucamides, FAGA) and combinations    thereof.-   41. The composition according to any of the preceding composition    paragraphs, wherein the composition further comprises one or more    enzymes selected from the group consisting of proteases, lipases,    cutinases, amylases, carbohydrases, cellulases, pectinases,    mannanases, arabinases, galactanases, xylanases and oxidases.-   42. The composition according to any of the preceding composition    paragraphs, wherein the enzyme is a protease, which is of animal,    vegetable or microbial origin.-   43. The composition according to any of the preceding composition    paragraphs, wherein the protease is chemically modified or protein    engineered.-   44. The composition according to any of the preceding composition    paragraphs, wherein the protease is a serine protease or a    metalloprotease, preferably an alkaline microbial protease or a    trypsin-like protease.-   45. The composition according to any of the preceding composition    paragraphs, wherein the protease is selected from the group    consisting of Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg,    subtilisin 309, subtilisin 147, subtilisin 168, trypsin of bovine    origin, trypsin of porcine origin and Fusarium protease.-   46. The composition according to any of the preceding composition    paragraphs, wherein the biofilm comprising components such as DNA or    PNAG is produced by any of the following; Acinetobacter sp.,    Aeromicrobium sp., Brevundimonas sp., Microbacterium sp.,    Micrococcus luteus, Pseudomonas sp., Staphylococcus sp., and    Stenotrophomonas sp. particularly, Bacillus subtillis, Escherichia    coli, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas    fluorescens, Yersinia pestis, Aggregatibacter actinomycetemcomitans,    Streptococcus pyogenes, Streptococcus dysgalactiae (group C strep),    Enterococcus faecalis, Listeria monocytogenes, Clostridium    difficile, Mycobacterium tuberculosis, Mycobacterium smegmatis;    Neisseria meningitides, Neisseria gonorrhea, nontypable Haemophilus    influenzae, Haemophilus ducreyi, Helicobacter pylon, Campylobacter    jejuni; Citrobacter rodentium, Salmonella enterica serovars typhi,    Salmonella typhimurium, Candida albicans, Aspergillus flavus,    Fusarium solani, and Cryptococcus neoformans, Acinetobacter    baumannii, Actinobacillus actinomycetemcomitans, Bdellovibrio    bacterivorous, Bordetella pertussis, Bordetella bronchiectasis,    Campylobacter jejuni, Comamonas denitrificans, Escherichia coli,    Haemophilus influenza, Klebsiella pneumoniae, Neisseria    meningitides, Pseudomonas aeruginosa, Shewanella oneidensis, Vibrio    cholera, Gram-positive bacteria, Bacillus licheniformis, Bacillus    subtilis, Enterococcus faecalis, Listeria monocytogenes, Micrococcus    luteus, Staphylococcus aureus, Staphylococcus epidermidis,    Staphylococcus haemolyticus, Streptococcus anginosus, Streptococcus    constellatus, Streptococcus salivarius, Staphylococcus lugdunesis,    Streptococcus intermedius, Streptococcus intermedius, Streptococcus    mutans, Streptococcus pneumoniae, Streptococcus pyogenes,    Aspergillus fumigatus and Candida albicans-   47. The composition according to any of the preceding composition    paragraphs, wherein the composition is a bar, a homogenous tablet, a    tablet having two or more layers, a pouch having one or more    compartments, a regular or compact powder, a granule, a paste, a    gel, or a regular, compact or concentrated liquid.-   48. The composition according to any of the preceding composition    paragraphs, wherein the composition is a cleaning composition    selected from liquid detergent, powder detergent and granule    detergent compositions.-   49. A method for laundering an item comprising the steps of:    -   a) Exposing an item to a wash liquor comprising an enzyme having        DNase activity and enzyme having hexosaminidase activity or a        composition according to any of paragraphs 22-48;    -   b) Completing at least one wash cycle; and    -   c) Optionally rinsing the item,-    wherein the item is a textile.-   50. Method according to paragraph 49, wherein the pH of the wash    liquor is in the range of 1 to 11.-   51. Method according to any of the preceding method paragraphs,    wherein the pH of the wash liquor is in the range 5.5 to 11, such as    in the range of 7 to 9, in the range of 7 to 8 or in the range of 7    to 8.5.-   52. Method according to any of the preceding method paragraphs,    wherein the temperature of the wash liquor is in the range of 5° C.    to 95° C., or in the range of 10° C. to 80° C., in the range of    10° C. to 70° C., in the range of 10° C. to 60° C., in the range of    10° C. to 50° C., in the range of 15° C. to 40° C., in the range of    20° C. to 40° C., in the range of 15° C. to 30° C. or in the range    of 20° C. to 30° C.-   53. Method according to any of the preceding method paragraphs,    wherein the temperature of the wash liquor is from about 20° C. to    about 40° C.-   54. Method according to any of the preceding method paragraphs,    wherein the temperature of the wash liquor is from about 15° C. to    about 30° C.-   55. Method according to any of the preceding method paragraphs,    wherein stains present on the item is pre-treated with an enzyme    having DNase activity and an enzyme having hexosaminidase activity    or a detergent composition according to any of paragraphs 22-48.-   56. Method according to any of the preceding method paragraphs,    wherein stickiness of the item is reduced.-   57. Method according to any of the preceding method paragraphs,    wherein redeposition of soil is reduced.-   58. Method according to any of the preceding method paragraphs,    wherein adherence of soil to the item is reduced or removed.-   59. Method according to any of the preceding method paragraphs,    wherein whiteness of the item is maintained or improved.-   60. Method according to any of the preceding method paragraphs,    wherein malodor is reduced or removed from the item.-   61. Method according to any of the preceding method paragraphs,    wherein the concentration of DNase in the wash liquor is preferably    at least at least 0.00001 mg/L or at least 0.0001 or at least 0.001    mg/L of enzyme or at least 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 2, 5,    10 or at least 15 μg active enzyme per liter of wash liquor,    optionally the concentration of enzyme in the wash liquor is in the    range of about 0.00002 mg/L to about 2 mg/L, about 0.0002 mg/L to    about 2 mg/L, about 0.002 mg/L to about 2 mg/L, such as about 0.02    mg/L to about 2 mg/L, such as about 0.2 mg/L to about 2 mg/L or in    the range of about 0.00001 mg/L to about 1 mg/L, about 0.0001 mg/L    to about 1 mg/L, about 0.0001 mg/L to about 10 mg/L or in the range    of in the range of about 0.001 mg/L to about 10 mg/L, or in the    range of about 0.01 mg/L to about 10 mg/L, or in in the range of    about 0.1 mg/L to about 10 mg/L per liter of wash liquor, and    optionally weight percent of composition of the DNase is present in    an amount corresponding to at least 0.0001 wt %, at least 0.0002 wt    %, at least 0.0005 wt %, preferably at least 0.001 wt %, preferably    at least 0.002 wt %, preferably at least 0.005 wt %, preferably at    least 0.01 wt %, preferably at least 0.02 wt %, preferably at least    0.05 wt % preferably at least 0.1 wt % of the total detergent    concentration, the concentration of DNase is preferably within the    ranges from about 0.0001 wt % to about 10 wt %, such as from about    0.0001 wt % to about 5 wt %, such as from about 0.0002 wt % to about    5 wt %, such as from about 0.001 wt % to about 1 wt %, such as from    about 0.005 wt % to about 1 wt %, such as from about 0.01 wt % to    about 1 wt %, such as from about 0.01 wt % to about 0.5 wt % or most    preferred from about 0.002 wt % to about 0.01 wt % active enzyme in    the total detergent concentration.-   62. Method according to any of the preceding method paragraphs,    wherein the concentration of the polypeptide having hexosaminidase    activity in the wash liquor is preferably at least at least 0.00001    mg/L or at least 0.0001 or at least 0.001 mg/L of enzyme or at least    0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 2, 5, 10 or at least 15 μg    active enzyme per liter of wash liquor, optionally the concentration    of enzyme in the wash liquor is in the range of about 0.00002 mg/L    to about 2 mg/L, about 0.0002 mg/L to about 2 mg/L, about 0.002 mg/L    to about 2 mg/L, such as about 0.02 mg/L to about 2 mg/L, such as    about 0.2 mg/L to about 2 mg/L or in the range of about 0.00001 mg/L    to about 1 mg/L, about 0.0001 mg/L to about 1 mg/L, about 0.0001    mg/L to about 10 mg/L or in the range of in the range of about 0.001    mg/L to about 10 mg/L, or in the range of about 0.01 mg/L to about    10 mg/L, or in in the range of about 0.1 mg/L to about 10 mg/L per    liter of wash liquor and optionally weight percent of the    hexosaminidase of the composition at least 0.0001 wt %, preferably    at least 0.001 wt %, preferably at least 0.002 wt %, preferably at    least 0.005 wt %, preferably at least 0.01 wt %, preferably at least    0.02 wt %, preferably at least 0.05 wt % preferably at least 0.1 wt    % of the total detergent concentration, preferably within the ranges    from about 0.0001 wt % to about 10 wt %, such as from about 0.001 wt    % to about 0.1 wt %, such as from about 0.005 wt % to about 0.1 wt    %, such as from about 0.01 wt % to about 0.1 wt %, such as from    about 0.01 wt % to about 0.5 wt % or most preferred from about 0.002    wt % to about 0.01 wt % active enzyme in the total detergent    concentration-   63. Item laundered according to the method of any of paragraphs    49-62.    Every maximum numerical limitation given throughout this    specification includes every lower numerical limitation, as if such    lower numerical limitations were expressly written herein. Every    minimum numerical limitation given throughout this specification    will include every higher numerical limitation, as if such higher    numerical limitations were expressly written herein. Every numerical    range given throughout this specification will include every    narrower numerical range that falls within such broader numerical    range, as if such narrower numerical ranges were all expressly    written herein.    Assays and Detergent Compositions    Detergent Compositions

The below mentioned detergent compositions may be used in combinationwith the enzyme of the invention.

Biotex Black (Liquid)

5-15% Anionic surfactants, <5% Nonionic surfactants, perfume, enzymes,DMDM and hydantoin.

Composition of Ariel Sensitive White & Color, Liquid DetergentComposition

Aqua, Alcohol Ethoxy Sulfate, Alcohol Ethoxylate, Amino Oxide, CitricAcid, C12-18 topped palm kernel fatty acid, Protease, Glycosidase,Amylase, Ethanol, 1,2 Propanediol, Sodium Formate, Calcium Chloride,Sodium hydroxide, Silicone Emulsion, Trans-sulphated EHDQ (theingredients are listed in descending order).

Composition of WFK IEC-A Model Detergent (Powder)

Ingredients: Linear sodium alkyl benzene sulfonate 8.8%, Ethoxylatedfatty alcohol C12-18 (7 EO) 4.7%, Sodium soap 3.2%, Anti foam DC2-4248S3.9%, Sodium aluminium silicate zeolite 4A 28.3%, Sodium carbonate11.6%, Sodium salt of a copolymer from acrylic and maleic acid (SokalanCP5) 2.4%, Sodium silicate 3.0%, Carboxymethylcellulose 1.2%, Dequest2066 2.8%, Optical whitener 0.2%, Sodium sulfate 6.5%, Protease 0.4%.

Composition of Model Detergent A (Liquid)

Ingredients: 12% LAS, 11% AEO Biosoft N25-7 (NI), 5% AEOS (SLES), 6% MPG(monopropylene glycol), 3% ethanol, 3% TEA, 2.75% coco soap, 2.75% soyasoap, 2% glycerol, 2% sodium hydroxide, 2% sodium citrate, 1% sodiumformate, 0.2% DTMPA and 0.2% PCA (all percentages are w/w)

Composition of Ariel Actilift (Liquid)

Ingredients: 5-15% Anionic surfactants; <5% Non-ionic surfactants,Phosphonates, Soap; Enzymes, Optical brighteners, Benzisothiazolinone,Methylisothiazolinone, Perfumes, Alpha-isomethyl ionone, Citronellol,Geraniol, Linalool.

Composition of Ariel Actilift Colour&Style (Liquid)

Ingredients: 5-15% Anionic surfactants; <5% Non-ionic surfactants,Phosphonates, Soap; Enzymes, Perfumes, Benzisothiazolinone,Methylisothiazolinone, Alpha-isomethyl ionone, Butylphenylmethylpropional, Citronellol, Geraniol, Linalool.

Composition of Persil Small & Mighty (Liquid)

Ingredients: 15-30% Anionic surfactants, Non-ionic surfacts, 5-15% Soap,<5% Polycarboxylates, Perfume, Phosphates, Optical Brighteners

Persil 2 in 1 with Comfort Passion Flower Powder

Sodium sulfate, Sodium carbonate, Sodium Dodecylbenzenesulfonate,Bentonite, Sodium Carbonate Peroxide, Sodium Silicate, Zeolite, Aqua,Citric acid, TAED, C12-15 Pareth-7, Stearic Acid, Parfum, Sodium AcrylicAcid/MA Copolymer, Cellulose Gum, Corn Starch Modified, Sodium chloride,Tetrasodium Etidronate, Calcium Sodium EDTMP, DisodiumAnilinomorpholinotriazinylaminostilbenesulfonate, Sodium bicarbonate,Phenylpropyl Ethyl Methicone, Butylphenyl Methylpropional, GlycerylStearates, Calcium carbonate, Sodium Polyacrylate, Alpha-IsomethylIonone, Disodium Distyrylbiphenyl Disulfonate, Cellulose, Protease,Limonene, PEG-75, Titanium dioxide, Dextrin, Sucrose, Sodium PolyarylSulphonate, CI 12490, CI 45100, CI 42090, Sodium Thiosulfate, CI 61585.

Persil Biological Powder

Sucrose, Sorbitol, Aluminum Silicate, Polyoxymethylene Melamine, SodiumPolyaryl Sulphonate, CI 61585, CI 45100, Lipase, Amylase, Xanthan gum,Hydroxypropyl methyl cellulose, CI 12490, Disodium DistyrylbiphenylDisulfonate, Sodium Thiosulfate, CI 42090, Mannanase, CI 11680,Etidronic Acid, Tetrasodium EDTA.

Persil Biological Tablets

Sodium carbonate, Sodium Carbonate Peroxide, Sodium bicarbonate,Zeolite, Aqua, Sodium Silicate, Sodium Lauryl Sulfate, Cellulose, TAED,Sodium Dodecylbenzenesulfonate, Hemicellulose, Lignin, Lauryl Glucoside,Sodium Acrylic Acid/MA Copolymer, Bentonite, Sodium chloride, Parfum,Tetrasodium Etidronate, Sodium sulfate, Sodium Polyacrylate,Dimethicone, Disodium Anilinomorpholinotriazinylaminostilbenesulfonate,Dodecylbenzene Sulfonic Acid, Trimethylsiloxysilicate, Calciumcarbonate, Cellulose, PEG-75, Titanium dioxide, Dextrin, Protease, CornStarch Modified, Sucrose, CI 12490, Sodium Polyaryl Sulphonate, SodiumThiosulfate, Amylase, Kaolin,

Persil Colour Care Biological Powder

Subtilisin, Imidazolidinone, Hexyl Cinnamal, Sucrose, Sorbitol, AluminumSilicate, Polyoxymethylene Melamine, CI 61585, CI 45100, Lipase,Amylase, Xanthan gum, Hydroxypropyl methyl cellulose, CI 12490, DisodiumDistyrylbiphenyl Disulfonate, Sodium Thiosulfate, CI 42090, Mannanase,CI 11680, Etidronic Acid, Tetrasodium EDTA.

Persil Colour Care Biological Tablets

Sodium bicarbonate, Sodium carbonate, Zeolite, Aqua, Sodium Silicate,Sodium Lauryl Sulfate, Cellulose Gum, Sodium Dodecylbenzenesulfonate,Lauryl Glucoside, Sodium chloride, Sodium Acrylic Acid/MA Copolymer,Parfum, Sodium Thioglycolate, PVP, Sodium sulfate, TetrasodiumEtidronate, Sodium Polyacrylate, Dimethicone, Bentonite, DodecylbenzeneSulfonic Acid, Trimethylsiloxysilicate, Calcium carbonate, Cellulose,PEG-75, Titanium dioxide, Dextrin, Protease, Corn Starch Modified,Sucrose, Sodium Thiosulfate, Amylase, CI 74160, Kaolin.

Persil Dual Action Capsules Bio

MEA-Dodecylbenzenesulfonate, MEA-Hydrogenated Cocoate, C12-15 Pareth-7,Dipropylene Glycol, Aqua, Tetrasodium Etidronate, Polyvinyl Alcohol,Glycerin, Aziridine, homopolymer ethoxylated, Propylene glycol, Parfum,Sodium Diethylenetriamine Pentamethylene Phosphonate, Sorbitol,MEA-Sulfate, Ethanolamine, Subtilisin, Glycol, ButylphenylMethylpropional, Boronic acid, (4-formylphenyl), Hexyl Cinnamal,Limonene, Linalool, Disodium Distyrylbiphenyl Disulfonate,Alpha-Isomethyl Ionone, Geraniol, Amylase, Polymeric Blue Colourant,Polymeric Yellow Colourant, Talc, Sodium chloride, Benzisothiazolinone,Mannanase, Denatonium Benzoate.

Persil 2 in 1 with Comfort Sunshiny Days Powder

Sodium sulfate, Sodium carbonate, Sodium Dodecylbenzenesulfonate,Bentonite, Sodium Carbonate Peroxide, Sodium Silicate, Zeolite, Aqua,Citric acid, TAED, C12-15 Pareth-7, Parfum, Stearic Acid, Sodium AcrylicAcid/MA Copolymer, Cellulose Gum, Corn Starch Modified, Sodium chloride,Tetrasodium Etidronate, Calcium Sodium EDTMP, DisodiumAnilinomorpholinotriazinylaminostilbenesulfonate, Sodium bicarbonate,Phenylpropyl Ethyl Methicone, Butylphenyl Methylpropional, GlycerylStearates, Calcium carbonate, Sodium Polyacrylate, Geraniol, DisodiumDistyrylbiphenyl Disulfonate, Cellulose, Protease, PEG-75, Titaniumdioxide, Dextrin, Sucrose, Sodium Polyaryl Sulphonate, CI 12490, CI45100, CI 42090, Sodium Thiosulfate, CI 61585.

Persil Small & Mighty 2in1 with Comfort Sunshiny Days

Aqua, C12-15 Pareth-7, Sodium Dodecylbenzenesulfonate, Propylene glycol,Sodium Hydrogenated Cocoate, Triethanolamine, Glycerin, TEA-HydrogenatedCocoate, Parfum, Sodium chloride, Polyquaternium-10, PVP, Polymeric PinkColourant, Sodium sulfate, Disodium Distyrylbiphenyl Disulfonate,Butylphenyl Methylpropional, Styrene/Acrylates Copolymer, HexylCinnamal, Citronellol, Eugenol, Polyvinyl Alcohol, Sodium acetate,Isopropyl alcohol, Polymeric Yellow Colourant, Sodium Lauryl Sulfate.

Persil Small & Mighty Bio

Aqua, MEA-Dodecylbenzenesulfonate, Propylene glycol, Sodium LaurethSulfate, C12-15 Pareth-7, TEA-Hydrogenated Cocoate, MEA-Citrate,Aziridine homopolymer ethoxylated, MEA-Etidronate, Triethanolamine,Parfum, Acrylates Copolymer, Sorbitol, MEA-Sulfate, Sodium Sulfite,Disodium Distyrylbiphenyl Disulfonate, Butylphenyl Methylpropional,Styrene/Acrylates Copolymer, Citronellol, Sodium sulfate, Peptides,salts, sugars from fermentation (process), Subtilisin, Glycerin, Boronicacid, (4-formylphenyl), Geraniol, Pectate Lyase, Amylase, Sodium LaurylSulfate, Mannanase, CI 42051.

Persil Small & Mighty Capsules Biological

MEA-Dodecylbenzenesulfonate, MEA-Hydrogenated Cocoate, C12-15 Pareth-7,Dipropylene Glycol, Aqua, Glycerin, Polyvinyl Alcohol, Parfum, Aziridinehomopolymer ethoxylated, Sodium Diethylenetriamine PentamethylenePhosphonate, Propylene glycol, Sorbitol, MEA-Sulfate, Ethanolamine,Subtilisin, Glycol, Butylphenyl Methylpropional, Hexyl Cinnamal, Starch,Boronic acid, (4-formylphenyl), Limonene, Linalool, DisodiumDistyrylbiphenyl Disulfonate, Alpha-Isomethyl Ionone, Geraniol, Amylase,Talc, Polymeric Blue Colourant, Sodium chloride, Benzisothiazolinone,Denatonium Benzoate, Polymeric Yellow Colourant, Mannanase.

Persil Small & Mighty Capsules Colour Care

MEA-Dodecylbenzenesulfonate, MEA-Hydrogenated Cocoate, C12-15 Pareth-7,Dipropylene Glycol, Aqua, Glycerin, Polyvinyl Alcohol, Parfum, Aziridinehomopolymer ethoxylated, Sodium Diethylenetriamine PentamethylenePhosphonate, Propylene glycol, MEA-Sulfate, Ethanolamine, PVP, Sorbitol,Butylphenyl Methylpropional, Subtilisin, Hexyl Cinnamal, Starch,Limonene, Linalool, Boronic acid, (4-formylphenyl), Alpha-IsomethylIonone, Geraniol, Talc, Polymeric Blue Colourant, Denatonium Benzoate,Polymeric Yellow Colourant.

Persil Small & Mighty Colour Care

Aqua, MEA-Dodecylbenzenesulfonate, Propylene glycol, Sodium LaurethSulfate, C12-15 Pareth-7, TEA-Hydrogenated Cocoate, MEA-Citrate,Aziridine homopolymer ethoxylated, MEA-Etidronate, Triethanolamine,Parfum, Acrylates Copolymer, Sorbitol, MEA-Sulfate, Sodium Sulfite,Glycerin, Butylphenyl Methylpropional, Citronellol, Sodium sulfate,Peptides, salts, sugars from fermentation (process), Styrene/AcrylatesCopolymer, Subtilisin, Boronic acid, (4-formylphenyl), Geraniol, PectateLyase, Amylase, Sodium Lauryl Sulfate, Mannanase, CI 61585, CI 45100.

Composition of Fairy Non-Bio (Liquid)

Ingredients: 15-30% Anionic Surfactants, 5-15% Non-Ionic Surfactants,Soap, Benzisothiazolinone, Methylisothiazolinone, Perfumes

Composition of Model Detergent T (Powder)

Ingredients: 11% LAS, 2% AS/AEOS, 2% soap, 3% AEO, 15.15% sodiumcarbonate, 3% sodium silicate, 18.75% zeolite, 0.15% chelant, 2% sodiumcitrate, 1.65% AA/MA copolymer, 2.5% CMC and 0.5% SRP (all percentagesare w/w).

Composition of Model Detergent X (Powder)

Ingredients: 16.5% LAS, 15% zeolite, 12% sodium disilicate, 20% sodiumcarbonate, 1% sokalan, 35.5% sodium sulfate (all percentages are w/w).

Composition of Ariel Actilift Colour&Style (Powder)

Ingredients: 15-30% Anionic surfactants, <5% Non-ionic surfactants,Phosphonates, Polycarboxylates, Zeolites; Enzymes, Perfumes, Hexylcinnamal.

Composition of Ariel Actilift (Powder)

Ingredients: 5-15% Anionic surfactants, Oxygen-based bleaching agents,<5% Non-ionic surfactants, Phosphonates, Polycarboxylates, Zeolites,Optical brighteners, Enzymes, Perfumes, Butylphenyl Methylpropional,Coumarin, Hexyl Cinnamal

Composition of Persil Megaperls (Powder)

Ingredients: 15-30% of the following: anionic surfactants, oxygen-basedbleaching agent and zeolites, less than 5% of the following: non-ionicsurfactants, phosphonates, polycarboxylates, soap, Further ingredients:Perfumes, Hexyl cinnamal, Benzyl salicylate, Linalool, opticalbrighteners, Enzymes and Citronellol.

Gain Liquid, Original:

Ingredients: Water, Alcohol Ethoxysulfate, Diethylene Glycol, AlcoholEthoxylate, Ethanolamine, Linear Alkyl Benzene Sulfonate, Sodium FattyAcids, Polyethyleneimine Ethoxylate, Citric Acid, Borax, Sodium CumeneSulfonate, Propylene Glycol, DTPA, Disodium Diaminostilbene Disulfonate,Dipropylethyl Tetramine, Sodium Hydroxide, Sodium Formate, CalciumFormate, Dimethicone, Amylase, Protease, Liquitint™, Hydrogenated CastorOil, Fragrance

Tide Liquid, Original:

Ingredients: Linear alkylbenzene sulfonate, propylene glycol, citricacid, sodium hydroxide, borax, ethanolamine, ethanol, alcohol sulfate,polyethyleneimine ethoxylate, sodium fatty acids, diquaterniumethoxysulfate, protease, diethylene glycol, laureth-9,alkyldimethylamine oxide, fragrance, amylase, disodium diaminostilbenedisulfonate, DTPA, sodium formate, calcium formate, polyethylene glycol4000, mannanase, Liquitint™ Blue, dimethicone.

Liquid Tide, Free and Gentle:

Water, sodium alcoholethoxy sulfate, propylene glycol, borax, ethanol,linear alkylbenzene sulfonate sodium, salt, polyethyleneimineethoxylate, diethylene glycol, trans sulfated & ethoxylatedhexamethylene diamine, alcohol ethoxylate, linear alkylbenzenesulfonate, MEA salt, sodium formate, sodium alkyl sulfate, DTPA, amineoxide, calcium formate, disodium diaminostilbene, disulfonate, amylase,protease, dimethicone, benzisothiazolinone

Tide Coldwater Liquid, Fresh Scent:

Water, alcoholethoxy sulfate, linear alkylbenzene sulfonate, diethyleneglycol, propylene glycol, ethanolamine, citric acid, Borax, alcoholsulfate, sodium hydroxide, polyethyleneimine, ethoxylate, sodium fattyacids, ethanol, protease, Laureth-9, diquaternium ethoxysulfate,lauramine oxide, sodium cumene, sulfonate, fragrance, DTPA, amylase,disodium, diaminostilbene, disulfonate, sodium formate, disodiumdistyrylbiphenyl disulfonate, calcium formate, polyethylene glycol 4000,mannanase, pectinase, Liquitint™ Blue, dimethicone

Tide TOTALCARE™ Liquid, Cool Cotton:

Water, alcoholethoxy sulfate, propylene glycol, sodium fatty acids,laurtrimonium chloride, ethanol, sodium hydroxide, sodium cumenesulfonate, citric acid, ethanolamine, diethylene glycol, siliconepolyether, borax, fragrance, polyethyleneimine ethoxylate, protease,Laureth-9,

DTPA, polyacrylamide quaternium chloride, disodium diaminostilbenedisulfonate, sodium formate, Liquitint™ Orange, dipropylethyltetraamine, dimethicone, cellulase,

Liquid Tide Plus Bleach Alternative™, Vivid White and Bright, Originaland Clean Breeze:

Water, sodium alcoholethoxy sulfate, sodium alkyl sulfate, MEA citrate,linear alkylbenzene sulfonate, MEA salt, propylene glycol, diethyleneglycol, polyethyleneimine ethoxylate, ethanol, sodium fatty acids,ethanolamine, lauramine oxide, borax, Laureth-9, DTPA, sodium cumenesulfonate, sodium formate, calcium formate, linear alkylbenzenesulfonate, sodium salt, alcohol sulfate, sodium hydroxide, diquaterniumethoxysulfate, fragrance, amylase, protease, mannanase, pectinase,disodium diaminostilbene disulfonate, benzisothiazolinone, Liquitint™Blue, dimethicone, dipropylethyl tetraamine.

Liquid Tide HE, Original Scent:

Water, Sodium alcoholethoxy sulfate, MEA citrate, Sodium Alkyl Sulfate,alcohol ethoxylate, linear alkylbenzene sulfonate, MEA salt, sodiumfatty acids, polyethyleneimine ethoxylate, diethylene glycol, propyleneglycol, diquaternium ethoxysulfate, borax, polyethyleneimine, ethoxylatepropoxylate, ethanol, sodium cumene sulfonate, fragrance, DTPA, disodiumdiaminostilbene disulfonate, Mannanase, cellulase, amylase, sodiumformate, calcium formate, Lauramine oxide, Liquitint™ Blue,Dimethicone/polydimethyl silicone.

Tide TOTALCARE HE Liquid, Renewing Rain:

Water, alcoholethoxy sulfate, linear alkylbenzene sulfonate, alcoholethoxylate, citric acid, Ethanolamine, sodium fatty acids, diethyleneglycol, propylene glycol, sodium hydroxide, borax, polyethyleneimineethoxylate, silicone polyether, ethanol, protease, sodium cumenesulfonate, diquaternium ethoxysulfate, Laureth-9, fragrance, amylase,DTPA, disodium diaminostilbene disulfonate, disodium distyrylbiphenyldisulfonate, sodium formate, calcium formate, mannanase, Liquitint™Orange, dimethicone, polyacrylamide quaternium chloride, cellulase,dipropylethyl tetraamine.

Tide Liquid HE Free:

Water, alcoholethoxy sulfate, diethylene glycol, monoethanolaminecitrate, sodium formate, propylene glycol, linear alkylbenzenesulfonates, ethanolamine, ethanol, polyethyleneimine ethoxylate,amylase, benzisothiazolin, borax, calcium formate, citric acid,diethylenetriamine pentaacetate sodium, dimethicone, diquaterniumethoxysulfate, disodium diaminostilbene disulfonate, Laureth-9,mannanase, protease, sodium cumene sulfonate, sodium fatty acids.

Tide Coldwater HE Liquid, Fresh Scent:

Water, alcoholethoxy sulfate, MEA Citrate, alcohol sulfate, Alcoholethoxylate, Linear alkylbenzene sulfonate MEA, sodium fatty acids,polyethyleneimine ethoxylate, diethylene glycol, propylene glycol,diquaternium ethoxysulfate, borax, polyethyleneimine ethoxylatepropoxylate, ethanol, sodium cumene sulfonate, fragrance, DTPA, disodiumdiaminostilbene disulfonate, protease, mannanase, cellulase, amylase,sodium formate, calcium formate, lauramine oxide, Liquitint™ Blue,dimethicone.

Tide for Coldwater HE Free Liquid:

Water, sodium alcoholethoxy sulfate, MEA Citrate, Linear alkylbenzenesulfonate: sodium salt, Alcohol ethoxylate, Linear alkylbenzenesulfonate: MEA salt, sodium fatty acids, polyethyleneimine ethoxylate,diethylene glycol, propylene glycol, diquaternium ethoxysulfate, Borax,protease, polyethyleneimine ethoxylate propoxylate, ethanol, sodiumcumene sulfonate, Amylase, citric acid, DTPA, disodium diaminostilbenedisulfonate, sodium formate, calcium formate, dimethicone.

Tide Simply Clean & Fresh:

Water, alcohol ethoxylate sulfate, linear alkylbenzene sulfonateSodium/Mea salts, propylene glycol, diethylene glycol, sodium formate,ethanol, borax, sodium fatty acids, fragrance, lauramine oxide, DTPA,Polyethylene amine ethoxylate, calcium formate, disodium diaminostilbenedisulfonate, dimethicone, tetramine, Liquitint™ Blue.

Tide Pods, Ocean Mist, Mystic Forest, Spring Meadow:

Linear alkylbenzene sulfonates, C12-16 Pareth-9, propylene glycol,alcoholethoxy sulfate, water, polyethyleneimine ethoxylate, glycerine,fatty acid salts, PEG-136 polyvinyl acetate, ethylene Diamine disuccinicsalt, monoethanolamine citrate, sodium bisulfite, diethylenetriaminepentaacetate sodium, disodium distyrylbiphenyl disulfonate, calciumformate, mannanase, exyloglucanase, sodium formate, hydrogenated castoroil, natalase, dyes, termamyl, subtilisin, benzisothiazolin, perfume.

Tide to Go:

Deionized water, Dipropylene Glycol Butyl Ether, Sodium Alkyl Sulfate,Hydrogen Peroxide, Ethanol, Magnesium Sulfate, Alkyl Dimethyl AmineOxide, Citric Acid, Sodium Hydroxide, Trimethoxy Benzoic Acid,Fragrance.

Tide Stain Release Liquid:

Water, Alkyl Ethoxylate, Linear Alkylbenzenesulfonate, HydrogenPeroxide, Diquaternium Ethoxysulfate, Ethanolamine, DisodiumDistyrylbiphenyl Disulfonate, tetrabutyl Ethylidinebisphenol, F&DCYellow 3, Fragrance.

Tide Stain Release Powder:

Sodium percarbonate, sodium sulfate, sodium carbonate, sodiumaluminosilicate, nonanoyloxy benzene sulfonate, sodium polyacrylate,water, sodium alkylbenzenesulfonate, DTPA, polyethylene glycol, sodiumpalmitate, amylase, protease, modified starch, FD&C Blue 1, fragrance.

Tide Stain Release, Pre-Treater Spray:

Water, Alkyl Ethoxylate, MEA Borate, Linear Alkylbenzenesulfonate,Propylene Glycol, Diquaternium Ethoxysulfate, Calcium Chlorideenzyme,Protease, Ethanolamine, Benzoisothiazolinone, Amylase, Sodium Citrate,Sodium Hydroxide, Fragrance.

Tide to Go Stain Eraser:

Water, Alkyl Amine Oxide, Dipropylene Glycol Phenyl Ether, HydrogenPeroxide, Citric Acid, Ethylene Diamine Disuccinic Acid Sodium salt,Sodium Alkyl Sulfate, Fragrance.

Tide Boost with Oxi:

Sodium bicarbonate, sodium carbonate, sodium percarbonate, alcoholethoxylate, sodium chloride, maleic/acrylic copolymer, nonanoyloxybenzene sulfonate, sodium sulfate, colorant, diethylenetriaminepentaacetate sodium salt, hydrated aluminosilicate (zeolite),polyethylene glycol, sodium alkylbenzene sulfonate, sodium palmitate,starch, water, fragrance.

Tide Stain Release Boost Duo Pac:

Polyvinyl Alcohol pouch film, wherein there is packed a liquid part anda powder part:

Liquid Ingredients: Dipropylene Glycol, diquaternium Ethoxysulfate,Water, Glycerin, Liquitint™ Orange, Powder Ingredients: sodiumpercarbonate, nonanoyloxy benzene sulfonate, sodium carbonate, sodiumsulfate, sodium aluminosilicate, sodium polyacrylate, sodiumalkylbenzenesulfonate, maleic/acrylic copolymer, water, amylase,polyethylene glycol, sodium palmitate, modified starch, protease,glycerine, DTPA, fragrance.Tide Ultra Stain Release:

Water, sodium alcoholethoxy sulfate, linear alkyl benzene sulfonate,sodium/MEA salts, MEA citrate, propylene glycol, polyethyleneimineethoxylate, ethanol, diethylene glycol, polyethyleneiminepropoxyethoxylate, sodium fatty acids, protease, borax, sodium cumenesulfonate, DTPA, fragrance, amylase, disodium diaminostilbenedisulfonate, calcium formate, sodium formate, gluconase, dimethicone,Liquitint™ Blue, mannanase.

Ultra Tide with a Touch of Downy® Powdered Detergent, April Fresh/CleanBreeze/April Essence:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Bentonite, Water, Sodium Percarbonate, SodiumPolyacrylate, Silicate, Alkyl Sulfate, Nonanoyloxybenzenesulfonate,DTPA, Polyethylene Glycol 4000, Silicone, Ethoxylate, fragrance,Polyethylene Oxide, Palmitic Acid, Disodium Diaminostilbene Disulfonate,Protease, Liquitint™ Red, FD&C Blue 1, Cellulase.

Ultra Tide with a Touch of Downy Clean Breeze:

Water, sodium alcoholethoxy sulfate, MEA citrate, linear alkyl benzenesulfonate: sodium/MEA salts, propylene glycol, polyethyleneimineethoxylate, ethanol, diethylene glycol, polyethyleneimine,propoxyethoxylate, diquaternium ethoxysulfate, alcohol sulfate,dimethicone, fragrance, borax, sodium fatty acids, DTPA, protease,sodium bisulfite, disodium diaminostilbene disulfonate, amylase,gluconase, castor oil, calcium formate, MEA, styrene acrylate copolymer,sodium formate, Liquitint™ Blue.

Ultra Tide with Downy Sun Blossom:

Water, sodium alcoholethoxy sulfate, MEA citrate, linear alkyl benzenesulfonate: sodium/MEA salts, propylene glycol, ethanol, diethyleneglycol, polyethyleneimine propoxyethoxylate, polyethyleneimineethoxylate, alcohol sulfate, dimethicone, fragrance, borax, sodium fattyacids, DTPA, protease, sodium bisulfite, disodium diaminostilbenedisulfonate, amylase, castor oil, calcium formate, MEA, styrene acrylatecopolymer, propanaminium propanamide, gluconase, sodium formate,Liquitint™ Blue.

Ultra Tide with Downy April Fresh/Sweet Dreams:

Water, sodium alcoholethoxy sulfate, MEA citrate, linear alkyl benzenesulfonate: sodium/MEA salts, propylene glycol, polyethyleneimineethoxylate, ethanol, diethylene glycol, polyethyleneiminpropoxyethoxylate, diquaternium ethoxysulfate, alcohol sulfate,dimethicone, fragrance, borax, sodium fatty acids, DTPA, protease,sodium bisulfite, disodium diaminostilbene disulfonate, amylase,gluconase, castor oil, calcium formate, MEA, styrene acrylate copolymer,propanaminium propanamide, sodium formate, Liquitint™ Blue.

Ultra Tide Free Powdered Detergent:

Sodium Carbonate, Sodium Aluminosilicate, Alkyl Sulfate, Sodium Sulfate,Linear Alkylbenzene Sulfonate, Water, Sodium polyacrylate, Silicate,Ethoxylate, Sodium percarbonate, Polyethylene Glycol 4000, Protease,Disodium Diaminostilbene Disulfonate, Silicone, Cellulase.

Ultra Tide Powdered Detergent, Clean Breeze/Spring Lavender/mountainSpring:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Alkyl Sulfate, Sodium Percarbonate, Water,Sodium Polyacrylate, Silicate, Nonanoyloxybenzenesulfonate, Ethoxylate,Polyethylene Glycol 4000, Fragrance, DTPA, Disodium DiaminostilbeneDisulfonate, Palmitic Acid, Protease, Silicone, Cellulase.

Ultra Tide HE (high Efficiency) Powdered Detergent, Clean Breeze:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Water, Nonanoyloxybenzenesulfonate, AlkylSulfate, Sodium Polyacrylate, Silicate, Sodium Percarbonate, Ethoxylate,Polyethylene Glycol 4000, Fragrance, DTPA, Palmitic Acid, DisodiumDiaminostilbene Disulfonate, Protease, Silicone, Cellulase.

Ultra Tide Coldwater Powdered Detergent, Fresh Scent:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, SodiumPercarbonate, Alkyl Sulfate, Linear Alkylbenzene Sulfonate, Water,Nonanoyloxybenzenesulfonate, Sodium Polyacrylate, Silicate, Ethoxylate,Polyethylene Glycol 4000, DTPA, Fragrance, Natalase, Palmitic Acid,Protease, Disodium, Diaminostilbene Disulfonate, FD&C Blue 1, Silicone,Cellulase, Alkyl Ether Sulfate.

Ultra Tide with Bleach Powdered Detergent, Clean Breeze:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Sodium Percarbonate,Nonanoyloxybenzenesulfonate, Alkyl Sulfate, Water, Silicate, SodiumPolyacrylate, Ethoxylate, Polyethylene Glycol 4000, Fragrance, DTPA,Palmitic Acid, Protease, Disodium Diaminostilbene Disulfonate, Silicone,FD&C Blue 1, Cellulase, Alkyl Ether Sulfate.

Ultra Tide with Febreeze Freshness™ Powdered Detergent, Spring Renewal:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Sodium Percarbonate, Alkyl Sulfate, Water,Sodium Polyacrylate, Silicate, Nonanoyloxybenzenesulfonate, Ethoxylate,Polyethylene Glycol 4000, DTPA, Fragrance, Cellulase, Protease, DisodiumDiaminostilbene Disulfonate, Silicone, FD&C Blue 1.

Liquid Tide Plus with Febreeze Freshness—Sport HE Active Fresh:

Water, Sodium alcoholethoxy sulfate, MEA citrate, linear alkylbenzenesulfonate, sodium salt, linear alkylbenzene sulfonate: MEA salt, alcoholethoxylate, sodium fatty acids, propylene glycol, diethylene glycol,polyethyleneimine ethoxylate propoxylate, diquaternium ethoxysulfate,Ethanol, sodium cumene sulfonate, borax, fragrance, DTPA, Sodiumbisulfate, disodium diaminostilbene disulfonate, Mannanase, cellulase,amylase, sodium formate, calcium formate, Lauramine oxide, Liquitint™Blue, Dimethicone/polydimethyl silicone.

Tide Plus Febreeze Freshness Spring & Renewal:

Water, sodium alcoholethoxy sulfate, linear alkyl benzene sulfonate:sodium/MEA salts, MEA citrate, propylene glycol, polyethyleneimineethoxylate, fragrance, ethanol, diethylene glycol, polyethyleneiminepropoxyethoxylate, protease, alcohol sulfate, borax, sodium fatty acids,DTPA, disodium diaminostilbene disulfonate, MEA, mannanase, gluconase,sodium formate, dimethicone, Liquitint™ Blue, tetramine.

Liquid Tide Plus with Febreeze Freshness, Sport HE Victory Fresh:

Water, Sodium alcoholethoxy sulfate, MEA citrate, linear alkylbenzenesulfonate, sodium salt, linear alkylbenzene sulfonate: MEA salt, alcoholethoxylate, sodium fatty acids, propylene glycol, diethylene glycol,polyethyleneimine ethoxylate propoxylate, diquaternium ethoxysulfate,ethanol, sodium cumene sulfonate, borax, fragrance, DTPA, Sodiumbisulfate, disodium diaminostilbene disulfonate, Mannanase, cellulase,amylase, sodium formate, calcium formate, Lauramine oxide, Liquitint™Blue, Dimethicone/polydimethyl silicone.

Tide Vivid White+Bright Powder, Original:

Sodium Carbonate, Sodium Aluminosilicate, Sodium Sulfate, LinearAlkylbenzene Sulfonate, Sodium Percarbonate,Nonanoyloxybenzenesulfonate, Alkyl Sulfate, Water, Silicate, SodiumPolyacrylate Ethoxylate, Polyethylene Glycol 4000, Fragrance, DTPA,Palmitic Acid, Protease, Disodium Diaminostilbene Disulfonate, Silicone,FD&C Blue 1, Cellulase, Alkyl Ether Sulfate.

Wash Assays

Mini Launder-O-Meter (MiniLOM) Model Wash System

MiniLOM is a mini wash system in which washes are performed in 50 mltest tubes placed in a Stuart rotator. Each tube simulates one smallwashing machine and during an experiment, each will contain a solutionof a specific detergent/enzyme system to be tested along with the soiledand unsoiled fabrics it is tested on. Mechanical stress is achieved viarotation (typically 20 rpm), and the temperature is controlled byplacement of the rotator in a heating cabinet/room.

EXAMPLES

Assay I: Testing of Hexosaminidase Activity

The hexosaminidase activity of the polypeptides listed in the tablebelow was determined using 4-nitrophenyl N-acetyl-β-D-glucosaminide(Sigma-Aldrich) as substrate. The enzymatic reaction was performed intriplicate in a 96 well flat bottom polystyrene microtiter plate (ThermoScientific) with the following conditions: 50 mM 2-(N-morpholino)ethanesulfonic acid pH 6 buffer, 1.5 mg/ml 4-nitrophenylN-acetyl-β-D-glucosaminide and 10, 20 or 50 μg/ml purified enzyme samplein a total reaction volume of 100 μl. Blank samples without polypeptidewere run in parallel. The reactions were carried out at 37° C. in aThermomixer comfort (Eppendorf). After 10 minutes of incubation, 5 μl 1M NaOH was added to each reaction mixture to stop the enzymaticreaction. The absorbance was read at 405 nm using a POLARstar Omegaplate reader (BMG LABTECH) to estimate the formation of 4-nitrophenolateion released because of enzymatic hydrolysis of the 4-nitrophenylN-acetyl-β-D-glucosaminide substrate. The results are summarized intable 1 below. The table shows the average absorbance measured at 405 nmfor each reaction performed in triplicate. It is seen that theabsorbance is higher for the reaction carried out with all thepolypeptides listed in the table below compared to blank withoutpolypeptide which demonstrates that all the tested polypeptides exhibithexosaminidase activity.

TABLE 1 Hexosaminidase activity. ΔA405 nm Enzyme (A405 nm_(sample) −Enzyme concentration A405 nm A405 nm_(blank)) Blank  0 μg/ml 0.158 — SEQID NO 3 10 μg/ml 1.352 1.194 SEQ ID NO 4 10 μg/ml 1.161 1.003 SEQ ID NO5 10 μg/ml 0.332 0.174 SEQ ID NO 6 10 μg/ml 0.321 0.163 SEQ ID NO 7 10μg/ml 2.903 2.745 SEQ ID NO 8 10 μg/ml 0.582 0.424 SEQ ID NO 9 10 μg/ml0.938 0.780 SEQ ID NO 10 10 μg/ml 1.152 0.994 SEQ ID NO 11 20 μg/ml0.376 0.197 SEQ ID NO 12 50 μg/ml 1.978 1.820 SEQ ID NO 13 50 μg/ml1.715 1.557 SEQ ID NO 14 50 μg/ml 2.455 2.297Assay IITesting of DNase Activity

DNase activity was determined on DNase Test Agar with Methyl Green (BD,Franklin Lakes, N.J., USA), which was prepared according to the manualfrom the supplier. Briefly, 21 g of agar was dissolved in 500 ml waterand then autoclaved for 15 min at 121° C. Autoclaved agar was temperatedto 48° C. in a water bath, and 20 ml of agar was poured into petridishesand allowed to solidify by incubation overnight at room temperature. Onsolidified agar plates, 5 μl of enzyme solutions are added, and DNaseactivity is observed as colorless zones around the spotted enzymesolutions.

Assay IIa

DNase activity was determined by fluorescence usingfluorescence-quenched DNA oligonucleotide probe. This probe emits signalafter nuclease degradation according to the manual from the supplier(DNase alert kit, Integrated DNA Technology, Coralville, Iowa, USA).Briefly, 5 μl of the substrate was added to 95 μl of DNase. If thesignal was too high, further dilutions of DNase was done in the adequatebuffer. Kinetic curve was measured for 20 min at 22° C. using aClariostar microplate reader (536 nm excitation, 556 nm emission).

Assay III

Wash performance is expressed as a delta remission value (ΔRem). Afterwashing and rinsing the swatches were spread out flat and allowed to airdry at room temperature overnight. All washes are evaluated the dayafter the wash. Light reflectance evaluations of the swatches were doneusing a Macbeth Color Eye 7000 reflectance spectrophotometer with verysmall aperture. The measurements were made without UV in the incidentlight and remission at 460 nm was extracted. Measurements were made onunwashed and washed swatches. The test swatch to be measured was placedon top of another swatch of same type and color (twin swatch). With onlyone swatch of each kind per beaker, a swatch from a replicate wash wasused in this way. Remission values for individual swatches werecalculated by subtracting the remission value of the unwashed swatchfrom the remission value of the washed swatch. The total washperformance for each stained swatch set was calculated as the sum ofindividual ΔRem.

Calculating the enzyme effect is done by taking the measurements fromwashed swatches with enzymes and subtracting with the measurements fromwashed swatches without enzyme for each stain. The total enzymeperformance is calculated as the sum of individual ΔRem_(enzyme).

Example 1

Extraction of EPS (Extracellular Polymeric Substances) from Pseudomonasfluorescens

A Pseudomonas fluorescens isolate from Iceland was used as a modelmicroorganism in the present example. Pseudomonas fluorescens wasrestreaked on Tryptone Soya Agar (TSA) (pH 7.3) (CM0131; Oxoid Ltd,Basingstoke, UK) and incubated for 1 day at 23° C. The strain wasinoculated into 10 mL of TSB and the culture was incubated with shakingfor 16 hours at 23° C. The overnight culture was diluted (1:100) in 200ml M63 supplemented medium (15 mM (NH₄)2SO₄, 100 mM KH₂PO₄, 1.8 μMFeSO₄, 1 mM MgSO₄.7H₂O, 0.4% (w/v) glycerol, 0.2% (w/v) Casamino acidsand 0.0001% (w/v) Thiamine) added to a Corning® CelIBIND® 225 cm² AngledNeck Cell Culture Flask with Vent Cap and incubated statically for 5days at 23° C. The biofilm culture was subsequently transferred to four50 ml Falcon tubes and pelleted by centrifugation (10 min, 8000 g, 25°C.), and the supernatants were discarded completely. The residualpellets from each Falcon tube were resuspended in 0.450 ml 3M NaCl toextract the surface-associated EPS (extracellular polymeric substances)and pooled in one test tube (5 ml, Eppendorf). The suspension wascentrifuged at 5000 g for 10 min at 25° C. and the 1.8 ml supernatantwas transferred to a new test tube as EPS fraction and stored at −20° C.until further use (termed crude EPS).

Fluorescent Assay with WGA-Alexa Fluor® 488

50 ul aliquots of the crude EPS were spotted on two wells of a Nuncmicrotiter plate and incubated at room temperature for 45 min.Supernatant was then removed and 50 ul of PBS buffer was added to onewell and 50 ul containing 20 ppm of enzyme (SEQ ID NO 14) was added tothe other well. The plate was incubated 1 hour at 30° C. Next,supernatants were removed and 50 ul of 10 ug/ml of wheat germ agglutininWGA-Alexa fluor488 fluorescent conjugate (Thermo Fischer Scientific,#W11261) was added to the wells. Alexa Fluor® 488 WGA binds to sialicacid and N-acetylglucosaminyl residues. The plate was incubated at roomtemperature for 15 min. Samples were washed with 50 ul water and thefluorescence at □_(excitation)=495 nm and □_(emission)=520 nm wasmeasured with a SpectraMax M3 plate reader instrument. The measurementsobtained are listed below in table 2.

TABLE 2 Fluorescent measurements of crude EPS from P. fluorescencetreated with or without hexosaminidase (SEQ ID NO 14) stained withWGA-Alexa Fluor488 dye. Treatment P. fluorescens EPS control PBS 122.724hexosaminidase (SEQ ID NO 14) 13.361Results from the fluorescent measurements show that the crude EPS samplefrom P. fluorescence is stained with WGA-Alexa Fluor488 suggesting thatit contains poly N-acetyl-glucosamine (PNAG) and is sensitive tohexosaminidase hydrolysis.DNA Content Analysis by Agarose Gel Electrophoresis10 ul aliquot of the crude EPS was treated with and without DNase (2ppm, SEQ ID NO 2) and subsequently subjected to agarose gelelectrophoresis. The gel was stained with SyBrSafe (Invitrogen). In theEPS sample, a prominent band with a high molecular mass was detected inthe 1% agarose gel corresponding to DNA. In contrast, in the EPS sampletreated with DNase a prominent band with a small molecular mass wasdetected corresponding to small degradation fragments of DNA. Thepresent study indicates that the crude EPS sample from P. fluorescenscontained DNA and that it was degraded by DNase treatment.

Example 2

Synergistic Effect Between Hexosaminidase and DNase on Deep-Cleaning inLiquid Model Detergent

50 ul aliquots of the crude EPS described in Example 1 were spotted onsterile textile swatches (WFK20A) and incubated for 30 min at ambienttemperature. The swatches (sterile or with EPS) were placed in 50 mLtest tubes and 10 mL of wash liquor (15⁰ dH water with 0.2 g/L iron(III)oxide nanopowder (544884; Sigma-Aldrich) with 3.33 g/L liquid model Adetergent) and 0.2 ug/ml enzyme(s) was added to each tube. Washeswithout enzyme were included as controls. The test tubes were placed ina Stuart rotator and incubated for 1 hour at 37° C. at 20 rpm. The washliquor was then removed, and the swatches were rinsed twice with 15⁰ dHwater and dried on filter paper over night. The remission (REM^(460 nm))values were measured using a Macbeth Color-Eye 7000 (CE7000), and aredisplayed in table 3. Delta values (REM^(460 nm) (swatch washed withenzyme)−REM^(460 nm) (swatch washed without enzyme)) are also indicated.

TABLE 3 Synergistic effect between hexosaminidase with SEQ ID NO 14 andDNase with SEQ ID NO 2 in deep-cleaning in model A detergent. Enzymeconcen- ΔREM^(460 nm) tration REM^(460 nm) (REM^(460 nm) _(with enzyme)− Enzyme (μg/ml) values REM^(460 nm) _(without enzyme)) EPS, No enzyme 040.3 EPS, hexosamini- 0.2 54.3 14.1 dase (SEQ ID NO 14) EPS, DNase 0.255.6 15.4 (SEQ ID NO 2) EPS, hexosamini- 0.2 + 0.2 75.5 35.2 dase (SEQID NO 14) + DNase (SEQ ID NO 2)As seen in table 3, the wash performance of the enzyme cocktailcomprising hexosaminidase (SEQ ID NO 14) and DNase (SEQ ID NO 2)(ΔREM460 nm (cocktail)=35.2) significantly exceeds the sum of theperformances seen for of the individual enzymes (ΔREM460 nm (sum ofindividual enzyme treatments)=29.5), showing a synergetic effect betweenthe enzymes. This also suggests that the different EPS componentstargeted by these enzymes are part of complex macromolecular structures,which shield other matrix components from enzymatic hydrolysis.

Example 3

Synergistic Effect Between Hexosaminidase and DNase on Deep-Cleaning inLiquid Model Detergent

A Pseudomonas fluorescens isolate from Iceland was used as a modelmicroorganism in the present example. Pseudomonas fluorescens wasrestreaked on Tryptone Soya Agar (TSA) (pH 7.3) (CM0131; Oxoid Ltd,Basingstoke, UK) and incubated for 1 day at 23° C. A single colony wasinoculated into 10 mL of TSB and the culture was incubated for 16 hoursat 23° C. with shaking (Tetramax 1000 at 460 rpm). After propagation,the culture was diluted to an OD600 of 0.03 in fresh TSB and 1.65 mLaliquots were added to the wells of 12-well polystyrene flat-bottommicroplates (3512; Costar, Corning Incorporated, Corning, N.Y., USA), inwhich round swatches (diameter 2 cm) of sterile textile (WFK20A) hadbeen placed. Sterile TSB was added to control wells. After 48 h at 23°C. (static incubation), the swatches were rinsed twice with 0.9% (w/v)NaCl.

Five rinsed swatches (sterile or with P. fluourescens) were placed in 50mL test tubes and 10 mL of wash liquor (15⁰ dH water with 0.2 g/Liron(III) oxide nanopowder (544884; Sigma-Aldrich) with 3.33 g/L liquidmodel A detergent) and 0.2 ppm enzymes was added to each tube. Washeswithout enzyme were included as controls. The test tubes were placed ina Stuart rotator and incubated for 1 hour at 30° C. at 20 rpm. The washliquor was then removed, and the swatches were rinsed twice with 15⁰ dHwater and dried on filter paper over night. The remission (REM^(460 nm))values were measured using a Macbeth Color-Eye 7000 (CE7000), and aredisplayed in table 4.

Delta values (REM^(460 nm) _((swatch washed with enzyme))−REM^(460 nm)_((swatch washed without enzyme))) are also indicated.

TABLE 4 Synergistic effect between hexosaminidase (SEQ ID NO 13) andDNase (SEQ ID NO 2) in deep-cleaning in model A detergent. Enzymeconcen- ΔREM^(460 nm) tration REM^(460 nm) (REM^(460 nm) _(with enzyme)− Enzyme (μg/ml) values REM^(460 nm) _(without enzyme)) Sterile wfk20A 070.6 swatch Biofilm swatch, no 0 42.6 enzyme Biofilm swatch, 0.2 45.02.4 hexosaminidase (SEQ ID NO 13) Biofilm swatch, 0.2 45.2 2.6 DNase(SEQ ID NO 2) Biofilm swatch, 0.2 + 0.2 55.0 12.3 hexosaminidase (SEQ IDNO 13) + DNase (SEQ ID NO 2)

As seen in table 4, an enzyme cocktail comprising hexosaminidase (SEQ IDNO 13) and DNase (SEQ ID NO 2) provides superior deep-cleaningproperties in model A detergent as compared to the individual enzymes.Given that the wash performance of the enzyme cocktail (ΔREM^(460 nm)(cocktail)=12.3) clearly exceeds the sum of the performances seen forthe individual enzymes (ΔREM^(460 nm) (sum of individual enzymetreatments)=5.0), this shows a significant synergetic effect between thetwo enzymes on the deep-cleaning properties in model A.

The invention claimed is:
 1. A method for laundering a textile,comprising: a. exposing a textile to a wash liquor comprising an enzymehaving DNase activity and an enzyme having hexosaminidase activity,wherein the enzyme having hexosaminidase activity has at least 80%sequence identity to SEQ ID NO: 19 or at least 85% sequence identity toSEQ ID NO: 20; and b. completing at least one wash cycle.
 2. The methodof claim 1, wherein the enzyme having hexosaminidase activity has atleast 85% sequence identity to SEQ ID NO:
 19. 3. The method of claim 1,wherein the enzyme having hexosaminidase activity has at least 90%sequence identity to SEQ ID NO:
 19. 4. The method of claim 1, whereinthe enzyme having hexosaminidase activity has at least 90% sequenceidentity to SEQ ID NO:
 20. 5. The method of claim 1, wherein the enzymehaving hexosaminidase activity has at least 95% sequence identity to SEQID NO:
 19. 6. The method of claim 1, wherein the enzyme havinghexosaminidase activity has at least 95% sequence identity to SEQ ID NO:20.
 7. The method of claim 1, wherein the enzyme having hexosaminidaseactivity comprises SEQ ID NO:
 19. 8. The method of claim 1, wherein theenzyme having hexosaminidase activity comprises SEQ ID NO:
 20. 9. Themethod of claim 1, wherein the enzyme having DNase activity has at least80% sequence identity to SEQ ID NO:
 2. 10. The method of claim 1,further comprising rinsing the textile.
 11. The method of claim 1,wherein the pH of the wash liquor is in the range 5.5 to
 11. 12. Themethod of claim 1, wherein the temperature of the wash liquor is fromabout 20° C. to about 40° C.
 13. The method of claim 1, wherein thetextile is pre-treated with an enzyme having DNase activity and anenzyme having hexosaminidase activity.
 14. The method of claim 1,wherein the concentration of DNase in the wash liquor is at least0.00001 mg/L of enzyme per liter of wash liquor.
 15. The method of claim1, wherein the concentration of the polypeptide having hexosaminidaseactivity in the wash liquor is at least 0.00001 mg/L of enzyme per literof wash liquor.